Library of biphenyl derivatives

ABSTRACT

This invention provides a library of biphenyl derivatives of formula I:  
                 
 
     wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , W, a, b and c are as defined in the specification, or a salt or stereoisomer thereof. The library is useful for identifying compounds having both β 2  adrenergic receptor agonist and muscarinic receptor antagonist activity.  
     Accordingly, this invention also provides methods of evaluating or screening a library of biphenyl derivatives to identifying compounds having such bifunctional activity.

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. ProvisionalApplication No. 60/447,843, filed on Feb. 14, 2003; and U.S. ProvisionalApplication No. 60/467,035, filed on May 1, 2003; the entire disclosuresof which are incorporated herein by reference in their entirety.

BACKGROUND OF THE INVENTION

[0002] 1. Field of the Invention

[0003] The present invention relates to a library or collection of novelbiphenyl derivatives. This invention also relates to methods of usingthe library of biphenyl derivatives to identify compounds having both β₂adrenergic receptor agonist and muscarinic receptor antagonist activity.Compounds having both β₂ adrenergic receptor agonist and muscarinicreceptor antagonist activity are useful for treating pulmonarydisorders.

[0004] 2. State of the Art

[0005] Pulmonary disorders, such as asthma and chronic obstructivepulmonary disease (COPD), are commonly treated with bronchodilators. Oneclass of bronchodilator in widespread use consists of β₂ adrenergicreceptor (adrenoceptor) agonists, such as albuterol, formoterol andsalmeterol. These compounds are generally administered by inhalation.Another class of bronchodilator consists of muscarinic receptorantagonists (anticholinergic compounds), such as ipratropium andtiotropium. These compounds are also typically administered byinhalation.

[0006] Pharmaceutical compositions containing both a β₂ adrenergicreceptor agonist and a muscarinic receptor antagonist are also known inthe art for use in treating pulmonary disorders. For example, U.S. Pat.No. 6,433,027 discloses medicament compositions containing a muscarinicreceptor antagonist, such as tiotropium bromide, and a β₂ adrenergicreceptor agonist, such as formoterol fumarate.

[0007] Although compounds having either β₂ adrenergic receptor agonistor muscarinic receptor antagonist activity are known, no compound havingboth β₂ adrenergic receptor agonist and muscarinic receptor antagonistactivity has been previously disclosed. Compounds possessing both β₂adrenergic receptor agonist and muscarinic receptor antagonist activityare highly desirable since such bifunctional compounds would providebronchodilation through two independent modes of action while havingsingle molecule pharmacokinetics. Accordingly, a need exists for methodsof identifying compounds having both β₂ adrenergic receptor agonist andmuscarinic receptor antagonist activity.

SUMMARY OF THE INVENTION

[0008] The present invention provides a library or collection of novelbiphenyl derivatives useful for identifying compounds having both β₂adrenergic receptor agonist and muscarinic receptor antagonist activity.This invention also provides methods of evaluating or screening such alibrary to identifying compounds having bifunctional activity.

[0009] Accordingly, in one of its composition aspects, the presentinvention is directed to a library comprising compounds of formula I:

[0010] wherein:

[0011] a is 0 or an integer of from 1 to 3;

[0012] each R¹ is independently selected from (1-4C)alkyl,(2-4C)alkenyl, (2-4C)alkynyl, (3-6C)cycloalkyl, cyano, halo, —OR^(1a),—C(O)OR^(1b), —SR^(1c), —S(O)R^(1d), —S(O)₂R^(1e) and —NR^(1f)R^(1g);

[0013] each of R^(1a), R^(1b), R^(1c), R^(1d), R^(1e), R^(1f) and R^(1g)is independently hydrogen, (1-4C)alkyl or phenyl-(1-4C)alkyl;

[0014] b is 0 or an integer of from 1 to 3;

[0015] each R² is independently selected from (1-4C)alkyl,(2-4C)alkenyl, (2-4C)alkynyl, (3-6C)cycloalkyl, cyano, halo, —OR^(2a),—C(O)OR^(2b), —SR^(2c), —S(O)R^(2d), —S(O)₂R^(2e) and —NR^(2f)R^(2g);

[0016] each of R^(2a), R^(2b), R^(2c), R^(2d), R^(2e), R^(2f) and R^(2g)is independently hydrogen, (1-4C)alkyl or phenyl-(1-4C)alkyl;

[0017] W is attached to the 3- or 4-position with respect to thenitrogen atom in the piperidine ring and represents O or NW^(a);

[0018] W^(a) is hydrogen or (1-4C)alkyl;

[0019] c is 0 or an integer of from 1 to 4;

[0020] each R³ is independently selected from (1-4C)alkyl,(2-4C)alkenyl, (2-4C)alkynyl, (3-6C)cycloalkyl, cyano, halo, —OR^(3a),—C(O)OR^(3b), —SR^(3c), —S(O)R^(3d), —S(O)₂R^(3e) and —NR^(3f)R^(3g); ortwo R³ groups are joined to form (1-3C)alkylene, (2-3C)alkenylene oroxiran-2,3-diyl;

[0021] each of R^(3a), R^(3b), R^(3c), R^(3d), R^(3e), R^(3f) and R^(3g)is independently hydrogen or (1-4C)alkyl;

[0022] R⁴ represents a divalent hydrocarbon group containing from 4 to28 carbon atoms and optionally containing from 1 to 10 heteroatomsselected independently from halo, oxygen, nitrogen and sulfur;

[0023] R⁵ represents hydrogen or (1-4C)alkyl;

[0024] R⁶ is —NR^(6a)CR^(6b)(O) or —CR^(6c)R^(6d)OR^(6e) and R⁷ ishydrogen; or R⁶ and R⁷ together form —NR^(7a)C(O)—CR^(7b)═CR^(7c)—,—CR^(7d)═CR^(7e)—C(O)—NR^(7f)—,—NR^(7g)C(O)—CR^(7h)R^(7i)—CR^(7j)R^(7k)— or—CR^(7l)R^(7m)—CR^(7n)R^(7o)—C(O)—NR^(7p)—;

[0025] each of R^(6a), R^(6b), R^(6c), R^(6d) and R^(6e) isindependently hydrogen or (1-4C)alkyl; and

[0026] each of R^(7a), R^(7b), R^(7c), R^(7d), R^(7e), R^(7f), R^(7g),R^(7h), R^(7i), R^(7j), R^(7k), R^(7l), R^(7m), R^(7n), R^(7o) andR^(7p) is independently hydrogen or (1-4C)alkyl;

[0027] or a salt or stereoisomer thereof.

[0028] In one embodiment, this invention is directed to a library thatcontains a compound of formula II:

[0029] wherein

[0030] R⁴ is as defined herein (including any specific or preferredembodiments);

[0031] W represents O or NH;

[0032] or a salt or stereoisomer thereof.

[0033] In another embodiment, this invention is directed to a librarythat contains a compound of formula III:

[0034] wherein

[0035] R⁴ is as defined herein (including any specific or preferredembodiments);

[0036] W represents O or NH;

[0037] or a salt or solvate or stereoisomer thereof.

[0038] In yet another embodiment, this invention is directed to alibrary that contains a compound of formula IV:

[0039] wherein

[0040] R⁴ is as defined herein (including any specific or preferredembodiments);

[0041] W represents O or NH;

[0042] or a salt or stereoisomer thereof.

[0043] The libraries or collections of this invention are useful foridentifying or finding or discovering compounds having both β₂adrenergic receptor agonist activity and muscarinic receptor antagonistactivity. Specifically, by evaluating or screening the members of thelibrary in the appropriate assays, those members having bifunctionalactivity, if any, can be identified.

[0044] Accordingly, in one of its method aspects, this inventionprovides a method of identifying a compound of formula I having both β₂adrenergic receptor agonist activity and muscarinic receptor antagonistactivity; the method comprising:

[0045] evaluating a library comprising compounds of formula I todetermine whether a compound in the library has both β₂ adrenergicreceptor agonist activity and muscarinic receptor antagonist activity.

[0046] In another of its method aspects, this invention provides amethod of identifying a compound of formula I having both β₂ adrenergicreceptor agonist activity and muscarinic receptor antagonist activity;the method comprising:

[0047] (a) preparing a library comprising compounds of formula I;

[0048] (b) evaluating the compounds of formula I in the library todetermine whether a compound has both β₂ adrenergic receptor agonistactivity and muscarinic receptor antagonist activity.

[0049] When identifying compounds having both β₂ adrenergic receptoragonist activity and muscarinic receptor antagonist activity, it isoften desirable to first identify compounds that bind to both a β₂adrenergic receptor and a muscarinic receptor. Accordingly, in anotherof its method aspects, this invention provides a method of identifying acompound of formula I that binds to a β₂ adrenergic receptor and amuscarinic receptor; the method comprising:

[0050] evaluating a library comprising compounds of formula I todetermine whether a compound in the library has a K_(i) value of lessthan about 300 nM for a β₂ adrenergic receptor and a K_(i) value lessthan 300 nM for a muscarinic receptor.

[0051] In still another of its method aspects, this invention provides amethod of identifying a compound of formula I that binds to a β₂adrenergic receptor and a muscarinic receptor; the method comprising:

[0052] (a) preparing a library comprising compounds of formula I;

[0053] (b) evaluating the compounds of formula I in the library todetermine whether a compound has a K_(i) value of less than about 300 nMfor a β₂ adrenergic receptor and a K_(i) value less than 300 nM for amuscarinic receptor.

[0054] In yet another of its method aspects, this invention provides amethod of identifying a compound of formula I that binds to a β₂adrenergic receptor and a muscarinic receptor; the method comprising:

[0055] evaluating a library comprising compounds of formula I todetermine whether a compound in the library has an EC₅₀ value of lessthan about 100 nM for a β₂ adrenergic receptor and a K_(i) value lessthan 100 nM for a muscarinic receptor.

[0056] In still another of its method aspects, this invention provides amethod of identifying a compound of formula I that binds to a β₂adrenergic receptor and a muscarinic receptor; the method comprising:

[0057] (a) preparing a library comprising compounds of formula I;

[0058] (b) evaluating the compounds of formula I in the library todetermine whether a compound has a EC₅₀ value of less than about 100 nMfor a β₂ adrenergic receptor and a K_(i) value less than 100 nM for amuscarinic receptor.

[0059] This invention is also directed to processes for preparing alibrary of compounds of formula I. Accordingly, in another of its methodaspects, this invention is directed to a process of preparing a librarycomprising compounds of formula I, the process comprising:

[0060] (a) reacting a compound of formula 1 or a salt thereof, with acompound of formula 2;

[0061] (b) reacting a compound of formula 3 or a salt thereof, with acompound of formula 4;

[0062] (c) coupling a compound of formula 5 with a compound of formula6;

[0063] (d) for a compound of formula I wherein R⁵ represents a hydrogenatom, reacting a compound of formula 3 with a compound of formula 7 or ahydrate thereof, in the presence of a reducing agent;

[0064] (e) reacting a compound of formula 1 with a compound of formula 8or a hydrate thereof, in the presence of a reducing agent;

[0065] (f) reacting a compound of formula 9, with a compound of formula10; or

[0066] (g) reacting a compound of formula 11 or a hydrate thereof, witha compound of formula 10, in the presence of a reducing agent; and

[0067] (h) removing any protecting groups to form a compound of formulaI; wherein the compounds of formula 1-11 are as defined therein;

[0068] (i) repeating step (a), (b), (c), (d), (e), (f) or (g) and step(h) to prepare a library comprising compounds of formula I.

DETAILED DESCRIPTION OF THE INVENTION

[0069] In one of its aspects, the present invention is directed to alibrary or collection comprising novel biphenyl derivatives useful foridentifying compounds having both β₂ adrenergic receptor agonist andmuscarinic receptor antagonist activity. In this regard, the compoundsof formula I which comprise the library contain one or more chiralcenters and therefore, this invention is directed a libraries containingracemic mixtures; pure stereoisomers (i.e., enantiomers ordiastereomers); stereoisomer-enriched mixtures and the like unlessotherwise indicated. When a particular stereoisomer is shown or namedherein, it will be understood by those skilled in the art that minoramounts of other stereoisomers may be present in the compositions ofthis invention unless otherwise indicated, provided that the utility ofthe composition as a whole is not eliminated by the presence of suchother isomers.

[0070] In particular, compounds of formula I contain a chiral center atthe carbon atom indicated by the symbol * in the following formula:

[0071] In one embodiment of this invention, the carbon atom identifiedby the symbol * has the (R) configuration. In this embodiment, it ispreferred for compounds of formula I to have the (R) configuration atthe carbon atom identified by the symbol * or to be enriched in astereoisomeric form having the (R) configuration at this carbon atom. Inanother embodiment of this invention, the carbon atom identified by thesymbol * has the (S) configuration. In this embodiment, it is preferredfor compounds of formula I to have the (S) configuration at the carbonatom identified by the symbol * or to be enriched in a stereoisomericform having the (S) configuration at this carbon atom. In some cases, inorder to optimize the β₂ adrenergic agonist activity of the compounds offormula I, it is preferred that the carbon atom identified by thesymbol * has the (R) configuration.

[0072] The compounds of formula I also contain several basic groups(e.g., amino groups) and therefore, the compounds of formula I can existas the free base or in various salt forms. All such salt forms areincluded within the scope of this invention. Furthermore, solvates ofcompounds of formula I or salts thereof are included within the scope ofthis invention.

[0073] Additionally, where applicable, all cis-trans or E/Z isomers(geometric isomers), tautomeric forms and topoisomeric forms of thecompounds of formula I are included within the scope of this inventionunless otherwise specified.

[0074] The nomenclature used herein to name the compounds of formula Iand intermediates thereof has generally been derived using thecommercially-available AutoNom software (MDL, San Leandro, Calif.).Typically, compounds of formula I wherein W is O have been named asester derivatives of biphenyl-2-ylcarbamic acid; and compounds offormula I wherein W is NW^(a) have been named as urea derivatives.

[0075] Representative Embodiments

[0076] The following substituents and values are intended to providerepresentative examples of various aspects and embodiments of thisinvention. These representative values are intended to further defineand illustrate such aspects and embodiments and are not intended toexclude other embodiments or to limit the scope of this invention. Inthis regard, the representation that a particular value or substituentis preferred is not intended in any way to exclude other values orsubstituents from this invention unless specifically indicated.

[0077] The libraries or collections of this invention are comprised ofcompounds of formula I or salts or stereoisomers thereof.

[0078] In particular embodiments of the compounds of formula I, a and bare independently 0, 1 or 2; including 0 or 1. In one embodiment, both aand b are 0.

[0079] When present, each R¹ may be at the 2, 3, 4, 5 or 6-position ofthe phenyl ring to which it is attached. In one embodiment, each R¹ isindependently selected from (1-4C)alkyl, halo, —OR^(1a) and—NR^(1f)R^(1g); such as methyl, fluoro, chloro, bromo, hydroxy, methoxy,amino, methylamino, dimethylamino and the like. Particular values for R¹are fluoro or chloro.

[0080] When present, each R² may be at the 3, 4, 5 or 6-position on thephenylene ring to which it is attached (where the carbon atom on thephenylene ring attached to the nitrogen atom is position 1). In oneembodiment, each R² is independently selected from (1-4C)alkyl, halo,—OR^(2a) and —NR^(2f)R^(2g); such as methyl, fluoro, chloro, bromo,hydroxy, methoxy, amino, methylamino, dimethylamino and the like.Particular values for R² are fluoro or chloro.

[0081] Each R^(1a), R^(1b), R^(1c), R^(1d), R^(1e), R^(1f) and R^(1g)and R^(2a), R^(2b), R^(2c), R^(2d), R^(2e), R^(2f) and R^(2g) as used inR¹ and R², respectively, is independently hydrogen, (1-4C)alkyl orphenyl-(1-4C)alkyl; such as hydrogen, methyl, ethyl, n-propyl,isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl and benzyl. In oneembodiment, these groups are independently hydrogen or (1-3C)alkyl. Inanother embodiment, these groups are independently hydrogen, methyl orethyl.

[0082] In one embodiment of this invention, W is O. In anotherembodiment, W is NW^(a).

[0083] Generally, it has been found that compounds in which W representsO exhibit particularly high affinity for muscarinic and β₂ adrenergicreceptors. Accordingly, in a particular embodiment of this invention, Wpreferably represents O.

[0084] When referring to W, particular mention may be made of compoundswherein W is attached to the piperidine ring at the 4-position withrespect to the nitrogen atom of the piperidine ring.

[0085] When W is NW^(a), W^(a) is hydrogen or (1-4C)alkyl; such ashydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl,isobutyl and tert-butyl. In one embodiment, W^(a) is hydrogen or(1-3C)alkyl. In another embodiment, W^(a) is hydrogen, methyl or ethyl;such as hydrogen or methyl. In yet another embodiment, W^(a) is hydrogenand NW^(a) is NH.

[0086] In a particular embodiment of the compounds of formula I, c is 0,1 or 2; including 0 or 1. In one embodiment, c is 0.

[0087] In one embodiment, each R³ is at the 3, 4 or 5-position on thepiperidine ring (where the nitrogen atom of the piperidine ring isposition 1). In another embodiment, R³ is at 4-position on thepiperidine ring. In a particular aspect of these embodiments, each R³ isindependently selected from (1-4C)alkyl; such as methyl, ethyl,n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl and tert-butyl. Inanother aspect, each R³ is independently methyl or ethyl.

[0088] In another embodiment, R³ is at the 1-position of the piperidinering, i.e., on the nitrogen atom of the piperidine ring thus forming aquaternary amine salt. In a particular aspect of this embodiment, eachR³ is independently selected from (1-4C)alkyl; such as methyl, ethyl,n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl and tert-butyl. Inanother aspect, each R³ is independently methyl or ethyl.

[0089] In yet another embodiment, two R³ groups are joined to form a(1-3C)alkylene or (2-3C)alkenylene group. For example, two R³ groups atthe 2 and 6-positions on the piperidine ring can be joined to form anethylene bridge (i.e., the piperidine ring and the R³ groups form an8-azabicyclo[3.2.1]octane ring); or two R³ groups at the 1 and4-positions on the piperidine ring can be joined to form an ethylenebridge (i.e., the piperidine ring and the R³ groups form an1-azabicyclo[2.2.2]octane ring). In this embodiment, other R³ groups asdefined herein may also be present.

[0090] In still another embodiment, two R³ groups are joined to form aoxiran-2,3-diyl group. For example, two R³ groups at the 2 and6-positions on the piperidine ring can be joined to form a3-oxatricyclo[3.3.1.0^(2,4)]nonane ring). In this embodiment, other R³groups as defined herein may also be present.

[0091] Each R^(3a), R^(3b), R^(3c), R^(3d), R^(3e), R^(3f) and R^(3g) asused in R³ is independently hydrogen or (1-4C)alkyl; such as hydrogen,methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl andtert-butyl. In one embodiment, these groups are independently hydrogenor (1-3C)alkyl. In another embodiment, these groups are independentlyhydrogen, methyl or ethyl.

[0092] In one embodiment of the compounds of formula I, R⁵ is hydrogenor (1-4C)alkyl; such as hydrogen, methyl, ethyl, n-propyl, isopropyl,n-butyl, sec-butyl, isobutyl and tert-butyl. In another embodiment, eachR⁵ is independently hydrogen, methyl or ethyl. In a particularembodiment, R⁵ is hydrogen.

[0093] In one embodiment of this invention, R⁶ is —NR^(6a)CR^(6b)(O) andR⁷ is hydrogen, where each of R^(6a) and R^(6b) is independentlyhydrogen or (1-4C)alkyl, such as hydrogen, methyl, ethyl, n-propyl,isopropyl, n-butyl, sec-butyl, isobutyl and tert-butyl. In oneembodiment, these groups are independently hydrogen or (1-3C)alkyl. Inanother embodiment, these groups are independently hydrogen, methyl orethyl. A particular value for R⁶ in this embodiment is —NHCHO.

[0094] In another embodiment, R⁶ and R⁷ together form—NR^(7a)C(O)—CR^(7b)═CR ^(7c)—, —CR^(7d)═CR^(7e)—C(O)—NR^(7f)—,—NR^(7g)C(O)—CR^(7h)R^(7i)—CR^(7j)R^(7k)— or—CR^(7l)R^(7m)—CR^(7n)R^(7o)—C(O)—NR^(7p)—; where each of R^(7a),R^(7b), R^(7c), R^(7d), R^(7e), R^(7f), R^(7g), R^(7h), R^(7i), R^(7j),R^(7k), R^(7l), R^(7m), R^(7n), R^(7o) and R^(7p) is independentlyhydrogen or (1-4C)alkyl; such as hydrogen, methyl, ethyl, n-propyl,isopropyl, n-butyl, sec-butyl, isobutyl and tert-butyl. In oneembodiment, these groups are independently hydrogen or (1-3C)alkyl. Inanother embodiment, these groups are independently hydrogen, methyl orethyl. Particular values for R⁶ and R⁷ in this embodiment are R⁶ and R⁷together form —NHC(O)—CH═CH—, —CH═CH—C(O)—NH—, —CH₂—CH₂—C(O)NH— or—NHC(O)—CH₂—CH₂—; including where R⁶ and R⁷ together form —NHC(O)—CH═CH—or —CH═CH—C(O)—NH—; and in particular, where R⁶ and R⁷ together form—NHC(O)—CH═CH— (i.e., the nitrogen atom is attached at R⁶ and the carbonatom is attached at R⁷ to form, together with the hydroxyphenyl ring towhich R⁶ and R⁷ are attached, a 8-hydroxy-2-oxo-1,2-dihydroquinolin-5-ylgroup).

[0095] In the compounds of formula I, R⁴ is a divalent hydrocarbon groupcontaining from 4 to 28 carbon atoms and optionally containing from 1 to10 heteroatoms selected independently from halo, oxygen, nitrogen andsulfur. In one embodiment, this group contains from 4 to 24 carbonatoms, including from 6 to 20 carbon atoms, such as from 8 to 18 carbonatoms; and optionally contains from 1 to 8 heteroatoms, including from 1to 6 heteroatoms.

[0096] The divalent hydrocarbon may contain any arrangement of atomsincluding alkylene, cycloalkylene, arylene, heteroarylene andheterocyclene groups or combinations thereof. The hydrocarbon group maybe interrupted by one or more heteroatoms or combinations of heteroatomsand carbon atoms to form various functional groups, such as ethers,thioethers, amines, amides, esters, carbamates, ureas, sulfones,sulfoxides, sulfonamides and the like.

[0097] In a particular aspect of this invention, the divalenthydrocarbon group of the compounds of formula I is a divalent group ofthe formula:

—(R^(4a))_(d)—(A¹)_(e)—(R^(4b))_(f)—Q—(R^(4c))_(g)—(A²)_(h)—(R^(d))_(i)—

[0098] wherein

[0099] d, e, f, g, h and i are each independently selected from 0 and 1;

[0100] R^(4a), R^(4b), R^(4c) and R^(4d) are each independently selectedfrom (1-10C)alkylene, (2-10C)alkenylene and (2-10C)alkynylene, whereineach alkylene, alkenylene or alkynylene group is unsubstituted orsubstituted with from 1 to 5 substituents independently selected from(1-4C)alkyl, fluoro, hydroxy, phenyl and phenyl-(1-4C)alkyl;

[0101] A¹ and A² are each independently selected from(3-7C)cycloalkylene, (6-10C)arylene, —O—(6-10C)arylene,(6-10C)arylene-O—, (2-9C)heteroarylene, —O-(2-9C)heteroarylene,(2-9C)heteroarylene-O— and (3-6C)heterocyclene, wherein eachcycloalkylene is unsubstituted or substituted with from 1 to 4substitutents selected independently from (1-4C)alkyl, and each arylene,heteroarylene or heterocyclene group is unsubstituted or substitutedwith from 1 to 4 substituents independently selected from halo,(1-4C)alkyl, (1-4C)alkoxy, —S-(1-4C)alkyl, —S(O)-(1-4C)alkyl,—S(O)₂-(1-4C)alkyl, —C(O)O(1-4C)alkyl, carboxy, cyano, hydroxy, nitro,trifluoromethyl and trifluoromethoxy;

[0102] Q is selected from a bond, —O—, —C(O)O—, —OC(O)—, —S—, —S(O)—,—S(O)₂—, —N(Q^(a))C(O)—, —C(O)N(Q^(b))—, —N(Q^(c))S(O)₂—,—S(O)₂N(Q^(d))—, —N(Q^(e))C(O)N(Q^(f))—, —N(Q^(g))S(O)₂N(Q^(h))—,—OC(O)N(Q^(i))—, —N(O^(j))C(O)O— and —N(Q^(k));

[0103] Q^(a), Q^(b), Q^(c), Q^(d), Q^(e), Q^(f), Q^(g), Q^(h), Q^(i),Q^(j) and Q^(k) are each independently selected from hydrogen,(1-6C)alkyl, A³ and (1-4C)alkylene-A⁴, wherein the alkyl group isunsubstituted or substituted with from 1 to 3 substituents independentlyselected from fluoro, hydroxy and (1-4C)alkoxy; or together with thenitrogen atom and the group R^(4b) or R^(4c) to which they are attached,form a 4-6 membered azacycloalkylene group;

[0104] A³ and A⁴ are each independently selected from (3-6C)cycloalkyl,(6-10C)aryl, (2-9C)heteroaryl and (3-6C)heterocyclyl, wherein eachcycloalkyl is unsubstituted or substituted with from 1 to 4substitutents selected independently from (1-4C)alkyl and each aryl,heteroaryl or heterocyclyl group is unsubstituted or substituted withfrom 1 to 4 substituents independently selected from halo, (1-4C)alkyland (1-4C)alkoxy.

[0105] In a particular embodiment, the values of each of the componentsR^(4a), A¹, R^(4b), Q, R^(4c), A² and R^(4d) are selected such that thenumber of contiguous atoms in the shortest chain between the twonitrogen atoms to which R⁴ is attached is in the range of from 4 to 16,(specifically, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16);including 8, 9, 10, 11, 12, 13 or 14; such as 8, 9, 10 or 11; or 9 or10. When selecting values for each variable in R⁴, it will beappreciated by those skilled in the art that values should be selectedsuch that a chemically stable group is formed.

[0106] When determining the number of contiguous atoms in the shortestchain between the two nitrogen atoms to which R⁴ is attached, eachcontiguous atom of the chain is counted consecutively starting from thefirst atom in the R⁴ group adjacent to the nitrogen of the piperidinering ending with the last atom in the R⁴ group adjacent to the nitrogenof the aminohydroxyethyl group. Where two or more chains are possible,the shortest chain is used to determine the number of contiguous atoms.As shown below, for example, when R⁴ is—(CH₂)₂—NHC(O)—CH₂-(phen-1,4-ylene)-CH₂—, there are 10 contiguous atomsin the shortest chain counted consecutively starting from the first atomin the R⁴ group adjacent to the nitrogen of the piperidine ring endingwith the last atom in the R⁴ group adjacent to the nitrogen of theaminohydroxyethyl group as shown below:

[0107] In one embodiment of R⁴, R^(4a) is selected from (1-10C)alkylene,(2-10C)alkenylene and (2-10C)alkynylene wherein the alkylene group isunsubstituted or substituted with 1 or 2 substituents independentlyselected from (1-4C)alkyl, hydroxy and phenyl. Representative examplesof particular values for R^(4a) are —(CH₂)₂—, —(CH₂)₃—, —(CH₂)₄—,—(CH₂)₅—, —(CH₂)₆—, —(CH₂)₇—, —(CH₂)₈—, —(CH₂)₉—, —(CH₂)₁₀—,—(CH₂)CH(CH₃)—, —(CH₂)C(CH₃)₂—, and —(CH₂)₂C(phenyl)₂-. In anotheraspect, R^(4a) is —(CH₂)C(═CH₂)—.

[0108] In one embodiment, d is 1.

[0109] In one embodiment, A¹ is an optionally substituted(3-7C)cycloalkylene group; including a cyclohexylene group, such ascyclohex-1,4-ylene and cyclohex-1,3-ylene; and a cyclopentylene group,such as cyclopent-1,3-ylene.

[0110] In another embodiment, A¹ is an optionally substituted(6-10C)arylene group, including a phenylene group, such asphen-1,4-ylene, phen-1,3-ylene and phen-1,2-ylene; and a naphthylenegroup, such as naphth-1,4-ylene and napth-1,5-ylene.

[0111] In yet another embodiment, A¹ is an optionally substituted(2-9C)heteroarylene group, including a pyridylene group, such aspyrid-1,4-ylene; a furylene group, such as fur-2,5-ylene andfur-2,4-ylene; a thienylene group, such as thien-2,5-ylene andthien-2,4-ylene; and a pyrrolylene, such as pyrrol-2,5-ylene andpyrrol-2,4-ylene.

[0112] In still another embodiment, A¹ is an optionally substituted(3-6C)heterocyclene group, including a piperidinylene group, such aspiperidin-1,4-ylene; and a pyrrolidinylene group, such aspyrrolidin-2,5-ylene.

[0113] In a particular embodiment, A¹ is an optionally substitutedphenylene, thienylene, cyclopentylene, cyclohexylene or piperidinylene.

[0114] In one embodiment, e is 0.

[0115] In a particular embodiment, R^(4b) is (1-5C)alkylene.Representative examples of particular values for R^(4b) are —CH₂—,—(CH₂)₂—, —(CH₂)₃—, —(CH₂)₄—, —(CH₂)₅—; including methylene, ethyleneand propylene.

[0116] In one embodiment, f is 0.

[0117] In a particular embodiment, Q is selected from a bond,—N(Q^(a))C(O)—, —C(O)N(Q^(b))—, —N(Q^(c))S(O)₂—, —S(O)₂N(Q^(d))—,—N(Q^(e))C(O)N(Q^(f))—, —OC(O)N(Q^(i))—, —N(Q^(j))C(O)O— or —N(Q^(k));such as where Q is a bond, —N(Q^(a))C(O)— or —C(O)N(Q^(b))—.Representative examples of particular values for Q are a bond, O, NH,—C(O)NH—, —C(O)N(CH₃)—, —NHC(O)—, —N(CH₃)C(O)—, —S(O)₂NH—,—S(O)₂N(CH₃)—, —NHS(O)₂—, —N(CH₃)S(O)₂— and —NHC(O)NH—. Another exampleof a value for Q, together with R^(4c), is —C(O)(piperidin-1,4-ylene).

[0118] In one embodiment, Q^(a), Q^(b), Q^(c), Q^(d), Q^(e), Q^(f),Q^(g), Q^(h), Q^(i), Q^(j) and Q^(k) are each independently selectedfrom hydrogen and (1-6C)alkyl, wherein the alkyl group is unsubstitutedor substituted with from 1 to 3 substituents independently selected fromfluoro, hydroxy and (1-4C)alkoxy. For example, Q^(a), Q^(b), Q^(c),Q^(d), Q^(e), Q^(f), Q^(g), Q^(h), Q^(i), Q^(j) and Q^(k) are eachindependently selected from hydrogen, and (1-3C)alkyl, includinghydrogen, methyl, ethyl, n-propyl and isopropyl. An example of a valuefor each of Q^(a), Q^(b), Q^(c), Q^(d), Q^(e), Q^(f), Q^(g), Q^(h),Q^(i), Q^(j) and Q^(k) is hydrogen.

[0119] In another embodiment, Q^(a), Q^(b), Q^(c), Q^(d), Q^(e), Q^(f),Q^(g), Q^(h), Q^(i), Q^(j) and Q^(k) together with the nitrogen atom andthe group R^(4b) or R^(4c) to which they are attached, form a 4-6membered azacycloalkylene group. For example, Q^(a) and Q^(b) togetherwith the nitrogen atom and the group R^(4b) or R^(4c) to which they areattached, form a piperidin-4-ylene group. By way of illustration, when Qrepresents —N(Q^(a))C(O)— and Q^(a) together with the nitrogen atom andthe group R^(4b) to which it is attached, forms a piperidin-4-ylenegroup, R⁴ is a group of formula:

[0120] Similarly, when Q represents —C(O)N(Q^(b))— and Q^(b) togetherwith the nitrogen atom and the group R^(4c) to which it is attached,forms a piperidin-4-ylene group, R⁴ is a group of formula:

[0121] In a particular embodiment, R^(4c) is (1-5C)alkylene.Representative examples of particular values for R^(4c) are —CH₂—,—(CH₂)₂—, —(CH₂)₃—, —(CH₂)₄—, —(CH₂)₅—; including methylene, ethyleneand propylene.

[0122] In one embodiment, A² is an optionally substituted(3-7C)cycloalkylene group; including a cyclohexylene group, such ascyclohex-1,4-ylene and cyclohex-1,3-ylene; and a cyclopentylene group,such as cyclopent-1,3-ylene.

[0123] In another embodiment, A² is an optionally substituted(6-10C)arylene group, including a phenylene group, such asphen-1,4-ylene, phen-1,3-ylene and phen-1,2-ylene; and a naphthylenegroup, such as naphth-1,4-ylene and napth-1,5-ylene.

[0124] In yet another embodiment, A² is an optionally substituted(2-9C)heteroarylene group, including a pyridylene group, such aspyrid-1,4-ylene; a furylene group, such as fur-2,5-ylene andfur-2,4-ylene; a thienylene group, such as thien-2,5-ylene andthien-2,4-ylene; and a pyrrolylene, such as pyrrol-2,5-ylene andpyrrol-2,4-ylene.

[0125] In still another embodiment, A² is an optionally substituted(3-6C)heterocyclene group, including a piperidinylene group, such aspiperidin-1,4-ylene; and a pyrrolidinylene group, such aspyrrolidin-2,5-ylene.

[0126] In a particular embodiment, A² is optionally substitutedphenylene, thienylene, cyclopentylene, cyclohexylene or piperidinylene.

[0127] By way of illustration, either A¹ or A² or both can be phenylene,such as phen-1,4-ylene or phen-1,3-ylene, where the phenylene group isunsubstituted or substituted with from 1 to 4 substituents independentlyselected from halo, (1-4C)alkyl, (1-4C)alkoxy, —S-(1-4C)alkyl,—S(O)-(1-4C)alkyl, —S(O)₂-(1-4C)alkyl, —C(O)O(1-4C)alkyl, carboxy,cyano, hydroxy, nitro, trifluoromethyl and trifluoromethoxy.Representative examples include phen-1,3-ylene, phen-1,4-ylene,4-chlorophen-1,3-ylene, 6-chlorophen-1,3-ylene, 4-methylphen-1,3-ylene,2-fluorophen-1,4-ylene, 2-chlorophen-1,4-ylene, 2-bromophen-1,4-ylene,2-iodophen-1,4-ylene, 2-methylphen-1,4-ylene, 2-methoxyphen-1,4-ylene,2-trifluoromethoxyphen-1,4-ylene, 3-nitrophen-1,4-ylene,3-chlorophen-1,4-ylene, 2,5-difluorophen-1,4-ylene,2,6-dichlorophen-1,4-ylene, 2,6-diiodophen-1,4-ylene,2-chloro-6-methylphen-1,4-ylene, 2-chloro-5-methoxyphen-1,4-ylene,2,3,5,6-tetrafluorophen-1,4-ylene.

[0128] Alternatively, A¹ or A² or both can be cyclopentylene orcyclohexylene; wherein the cyclopentylene or cyclohexylene group isunsubstituted or substituted with (1-4C)alkyl. Representative examplesinclude cis-cyclopent-1,3-ylene, trans-cyclopent-1,3-ylene,cis-cyclohex-1,4-ylene and trans-cyclohex-1,4-ylene. A¹ or A² or bothcan also be optionally substituted thienylene or piperidinylene, forexample, thien-2,5-ylene or piperidin-1,4-ylene.

[0129] In one embodiment, R^(4d) is selected from (1-10C)alkylene,(2-10C)alkenylene and (2-10C)alkynylene wherein the alkylene isunsubstituted or substituted with 1 or 2 substituents independentlyselected from (1-4C)alkyl, hydroxy and phenyl. Representative examplesof particular values for R^(4d) are —(CH₂)—, —(CH₂)₂—, —(CH₂)₃—,—(CH₂)₄—, —(CH₂)₅—, —(CH₂)₆—, —(CH₂)₇—, —(CH₂)₈—, —(CH₂)₉—, —(CH₂)10—and —(CH₂)CH(CH₃)—(CH₂)—C(CH₃)₂—(CH₂)₂—.

[0130] In a particular embodiment, R⁴ is a divalent group of theformula: —(R^(4a))_(d)— where R^(4a) is (4-10C)alkylene. In one aspectof this embodiment, R⁴ is a divalent group of the formula: —(CH₂)_(j)—where j is 8, 9 or 10. Examples of particular values for R⁴ in thisembodiment are —(CH₂)₄—, —(CH₂)₅—, —(CH₂)₆—, —(CH₂)₇—, —(CH₂)₈—,—(CH₂)₉, and —(CH₂)₁₀—; including —(CH₂)₈—, —(CH₂)₉, and —(CH₂)₁₀—.

[0131] In another particular embodiment, R⁴ is a divalent group of theformula:

—(R^(4a))_(d)—(A²)_(h)—(R^(4d))_(i)—

[0132] where R^(4a) is (1-10C)alkylene, such as —(CH₂)—, —(CH₂)₂—,—(CH₂)₃—; A² is (6-10C)arylene, such as phen-1,4-ylene orphen-1,3-ylene, or (2-9C)heteroarylene, such as thien-2,5-ylene orthien-2,4-ylene; and R^(4d) is (1-10C)alkylene, such as —(CH₂)—,—(CH₂)₂—, —(CH₂)₃—. Examples of particular values for R⁴ in thisembodiment are —CH₂)-(phen-1,4-ylene)-(CH₂)—;—(CH₂)-(phen-1,4-ylene)-(CH₂)₂—; —(CH₂)-(phen-1,4-ylene)-(CH₂)₃—;—(CH₂)₂-(phen-1,4-ylene)-(CH₂)—; —CH₂)₂-(phen-1,4-ylene)-(CH₂)₂—;—(CH₂)₂-(phen-1,4-ylene)-(CH₂)₃—; —(CH₂)₃-(phen-1,4-ylene)-(CH₂)—;—(CH₂)₃-(phen-1,4-ylene)-(CH₂)₂—, —(CH₂)₃-(phen-1,4-ylene)-(CH₂)₃—,—(CH₂)₄-(phen-1,4-ylene)-(CH₂)—; —(CH₂)₄-(phen-1,4-ylene)-(CH₂)₂— and—(CH₂)₄-(phen-1,4-ylene)-(CH₂)₃—.

[0133] In yet another particular embodiment, R⁴ is a divalent group ofthe formula:

—(R^(4a))_(d)—Q—(A²)_(h)—(R^(4d))_(i)—

[0134] where Q is —O— or —N(Q^(k))—; Q^(k) is hydrogen or (1-3C)alkyl,such as methyl or ethyl; R^(4a) is (1-10C)alkylene, such as —(CH₂)—,—(CH₂)₂—, —(CH₂)₃—; A² is (6-10C)arylene, such as phen-1,4-ylene orphen-1,3-ylene, or (2-9C)heteroarylene, such as thien-2,5-ylene orthien-2,4-ylene; and R^(4d) is (1-10C)alkylene, such as —(CH₂)—,—(CH₂)₂—, —(CH₂)₃—. Examples of particular values for R⁴ in thisembodiment are —(CH₂)₂—O-(phen-1,4-ylene)-(CH₂)—;—(CH₂)₂—O-(phen-1,4-ylene)-(CH₂)₂—; —(CH₂)₂—O-(phen-1,4-ylene)-(CH₂)₃—;—(CH₂)₃—O-(phen-1,4-ylene)-(CH₂)—; —(CH₂)₃—O-(phen-1,4-ylene)-(CH₂)₂—;—(CH₂)₃—O-(phen-1,4-ylene)-(CH₂)₃—; —(CH₂)₂—NH-(phen-1,4-ylene)-(CH₂)—;—(CH₂)₂—NH-(phen-1,4-ylene)-(CH₂)₂—;—(CH₂)₂—NH-(phen-1,4-ylene)-(CH₂)₃—; —(CH₂)₃—NH-(phen-1,4-ylene)-(CH₂)—;—(CH₂)₃—NH-(phen-1,4-ylene)-(CH₂)₂— and—(CH₂)₃—NH-(phen-1,4-ylene)-(CH₂)₃—.

[0135] In yet another particular embodiment, R⁴ is a divalent group ofthe formula:

—(R^(4a))_(d)—(A¹)_(e)—(R^(4b))_(f)—Q—(R^(4c))_(g)—(A²)_(h)—(R^(4d))_(i)—

[0136] where Q is —N(Q^(a))C(O)— or —C(O)N(Q^(b))—. A particular valuefor R⁴ in this embodiment is the formula:

[0137] where m is an integer from 2 to 10; and n is an integer from 2 to10; provided that m+n is an integer from 4 to 12. In this formula forR⁴, d and g are 1 and e, f, h and i are 0; and R^(4a) is —(CH₂)_(m)—,R^(4c) is —(CH₂)_(n)— and Q is —C(O)NH—. Particular values for m are 2or 3; and for n, 4, 5 or 6.

[0138] Another particular value for R⁴ is the formula:

[0139] where o is an integer from 2 to 7; and p is an integer from 1 to6; provided that o+p is an integer from 3 to 8. In this formula for R⁴,d, h and i are 1 and e, f and g are 0; and R^(4a) is —(CH₂)_(o)—, A² isphen-1,4-ylene, R^(4d) is —(CH₂)_(p)— and Q is —C(O)NH—. Particularvalues for o are 2 or 3; and for p, 1 or 2. In this embodiment, thephen-1,4-ylene group may be optionally substituted as defined herein forA².

[0140] Another particular value for R⁴ is the formula:

[0141] where q is an integer from 2 to 6; r is an integer from 1 to 5;and s is an integer from 1 to 5; provided that q+r+s is an integer from4 to 8. In this formula for R⁴, d, g, h and i are 1 and e and f are 0;and R^(4a) is —(CH₂)_(q)—, R^(4c) is —(CH₂)_(r)—, A² is 1,4-phenylene,R^(4d) is —(CH₂)_(s)— and Q is —C(O)NH—. Particular values for q are 2or 3; for r, 1 or 2; and for s, 1 or 2. In this embodiment, thephen-1,4-ylene group may be optionally substituted as defined herein forA².

[0142] Another particular value for R⁴ is the formula:

[0143] where t is an integer from 2 to 10; and u is an integer from 2 to10; provided that t+u is an integer from 4 to 12. In this formula forR⁴, d and g are 1 and e, f, h and i are 0; and R^(4a) is —(CH₂)_(t)—,R^(4c) is —(CH₂)_(u)— and Q is —NHC(O)—. Particular values for t are 2or 3; and for u, 4, 5 or 6.

[0144] Another particular value for R⁴ is the formula:

[0145] where v is an integer from 2 to 7; and w is an integer from 1 to6; provided that v+w is an integer from 3 to 8. In this formula for R⁴,d, h and i are 1 and e, f and g are 0; and R^(4a) is —(CH₂)_(v)—, A² is1,4-phenylene, R^(4d) is —(CH₂)_(w)— and Q is —NHC(O)—. Particularvalues for v are 2 or 3; and for w, 1 or 2. In this embodiment, thephen-1,4-ylene group may be optionally substituted as defined herein forA².

[0146] Another particular value for R⁴ is the formula:

[0147] where x is an integer from 2 to 6; y is an integer from 1 to 5;and z is an integer from 1 to 5; provided that x+y+z is an integer from4 to 8. In this formula for R⁴, d, g, h and i are 1 and e and f are 0;and R^(4a) is —(CH₂)_(x)—, R^(4c) is —(CH₂)_(y)—, A² is 1,4-phenylene,R^(4d) is —(CH₂)_(z)— and Q is —NHC(O)—. Particular values for x are 2or 3; for y, 1 or 2; and for z, 1 or 2. In this embodiment, thephen-1,4-ylene group may be optionally substituted as defined herein forA².

[0148] By way of further illustration, R⁴ can be selected from:

[0149] —(CH₂)₇—;

[0150] —(CH₂)₈—;

[0151] —(CH₂)₉—;

[0152] —(CH₂)₁₀—;

[0153] —(CH₂)₁₁—;

[0154] —(CH₂)₂C(O)NH(CH₂)₅—;

[0155] —(CH₂)₂N(CH₃)C(O)(CH₂)₅—;

[0156] —(CH₂)₂C(O)NH(phen-1,4-ylene)CH₂—;

[0157] —(CH₂)₂NHC(O)(phen-1,4-ylene)CH₂—;

[0158] —(CH₂)₂NHC(O)NH(CH₂)₅—;

[0159] —(CH₂)₃NHC(O)NH(CH₂)₅—;

[0160] —(CH₂)₂C(O)NHCH₂(cyclohex-1,3-ylene)CH₂—;

[0161] —(CH₂)₂NHC(O)(cyclopent-1,3-ylene)-;

[0162] —(CH₂)₂NHC(O)NH(phen-1,4-ylene)(CH₂)₂—;

[0163] 1-[—(CH₂)₂C(O)](piperidin-4-yl)(CH₂)₂—;

[0164] —(CH₂)₂NHC(O)(trans-cyclohex-1,4-ylene)CH₂—;

[0165] —(CH₂)₂NHC(O)(cis-cyclopent-1,3-ylene)-;

[0166] —(CH₂)₂NH(phen-1,4-ylene)(CH₂)₂—;

[0167] 1-[—(CH₂)₂NHC(O)](piperidin-4-yl)(CH₂)₂—;

[0168] —CH₂(phen-1,4-ylene)NH(phen-1,4-ylene)CH₂—;

[0169] —(CH₂)₂C(O)NHCH₂(phen-1,3-ylene)CH₂—;

[0170] —(CH₂)₂C(O)NHCH₂(pyrid-2,6-ylene)CH₂—;

[0171] —(CH₂)₂C(O)NH(cis-cyclohex-1,4-ylene)CH₂—;

[0172] —(CH₂)₂C(O)NH(trans-cyclohex-1,4-ylene)CH₂—;

[0173] —(CH₂)₂NHC(O)(cis-cyclopent-1,3-ylene)CH₂—;

[0174] —(CH₂)₂N(CH₃)C(O)(phen-1,3-ylene)CH₂—;

[0175] —(CH₂)₂N(CH₃)C(O)(trans-cyclohex-1,4-ylene)CH₂—;

[0176] —(CH₂)₂C(O)NH(phen-1,4-ylene)CH₂—;

[0177] —(CH₂)₂C(O)NH(phen-1,4-ylene)C*H(CH₃)—((S)-isomer);

[0178] —(CH₂)₂C(O)NH(phen-1,4-ylene)C*H(CH₃)—((R)-isomer);

[0179] 2-[(S)—(—CH₂—](pyrrolidin-1-yl)C(O)(CH₂)₄—;

[0180] 2-[(S)—(—CH₂—](pyrrolidin-1-yl)C(O)(phen-1,4-ylene)CH₂—;

[0181] —(CH₂)₂C(O)NH(4-chlorophen-1,3-ylene)CH₂—;

[0182] —CH₂(2-fluorophen-1,3-ylene)CH₂—;

[0183] —(CH₂)₂C(O)NH(4-methylphen-1,3-ylene)CH₂—;

[0184] —(CH₂)₂C(O)NH(6-chlorophen-1,3-ylene)CH₂—;

[0185] —(CH₂)₂C(O)NH(2-chlorophen-1,4-ylene)CH₂—;

[0186] —(CH₂)₂C(O)NH(2,6-dichlorophen-1,4-ylene)CH₂—;

[0187] —(CH₂)₂NHC(O)NHCH₂(phen-1,3-ylene)CH₂—;

[0188] 4-[—CH₂—](piperidin-1-yl)C(O)(phen-1,4-ylene)CH₂—;

[0189] —(CH₂)₂C(O)N(CH₂CH₃)(phen-1,4-ylene)CH₂—;

[0190] 1-[—(CH₂)₂NHC(O)](piperidin-4-yl)-;

[0191] —(CH₂)₂C(O)NH(phen-1,4-ylene)(CH₂)₂—;

[0192] —(CH₂)₂NHC(O)(thien-2,5-ylene)CH₂—;

[0193] —(CH₂)₂N(CH₃)C(O)(3-nitrophen-1,4-ylene)CH₂—;

[0194] —(CH₂)₂N(CH₃)C(O)(trans-cyclohex-1,4-ylene)-;

[0195] 1-[—CH₂(2-fluorophen-1,3-ylene)CH₂](piperidin-4-yl)-;

[0196] 5-[—(CH₂)₂NHC(O)](pyrid-2-yl)CH₂—;

[0197] —(CH₂)₂(phen-1,4-ylene)(CH₂)₂—;

[0198] —(CH₂)₃(thien-2,5-ylene)(CH₂)₃—;

[0199] —(CH₂)₂(phen-1,4-ylene)NH(phen-1,4-ylene)(CH₂)₂—;

[0200] —CH₂(phen-1,2-ylene)NH(phen-1,4-ylene)(CH₂)₂—;

[0201] 1-[—CH₂(2-fluorophen-1,3-ylene)CH₂](piperidin-4-yl)(CH₂)₂—;

[0202] 1-[—CH₂(2-fluorophen-1,3-ylene)CH₂](piperidin-4-yl)CH₂—;

[0203] —(CH₂)₂C(O)NH(3-chlorophen-1,4-ylene)CH₂—;

[0204] —(CH₂)₂C(O)NH(2-(CF₃O—)phen-1,4-ylene)CH₂—;

[0205] —(CH₂)₃(phen-1,3-ylene)NH(phen-1,4-ylene)(CH₂)₂—;

[0206] —(CH₂)₂S(O)₂NH(CH₂)₅—;

[0207] —CH₂(phen-1,3-ylene)NH(phen-1,4-ylene)(CH₂)₂—;

[0208] —(CH₂)₂C(O)NH(2-iodophen-1,4-ylene)CH₂—;

[0209] —(CH₂)₂C(O)NH(2-chloro-5-methoxyphen-1,4-ylene)CH₂—;

[0210] —(CH₂)₂C(O)NH(2-chloro-6-methylphen-1,4-ylene)CH₂—;

[0211] —(CH₂)₂C(O)NH(CH₂)₅—;

[0212] —(CH₂)₂N(CH₃)S(O)₂(phen-1,4-ylene)CH₂—;

[0213] —(CH₂)₂C(O)NH(2-bromophen-1,4-ylene)CH₂—;

[0214] —(CH₂)₃(phen-1,4-ylene)NH(phen-1,4-ylene)(CH₂)₂—;

[0215] —(CH₂)₃(phen-1,2-ylene)NH(phen-1,4-ylene)(CH₂)₂—;

[0216] 1-[—CH₂(2-fluorophen-1,3-ylene)CH₂](piperidin-4-yl)(CH₂)₃—;

[0217] —(CH₂)₂C(O)NH(2-methoxyphen-1,4-ylene)CH₂—;

[0218] —(CH₂)₅NH(phen-1,4-ylene)(CH₂)₂—;

[0219] 4-[—(CH₂)₂-](piperidin-1-yl)(phen-1,4-ylene)(CH₂)₂—;

[0220] —(CH₂)₂C(O)NH(phen-1,4-ylene)CH(CH₃)CH₂—;

[0221] —(CH₂)₂-(trans-cyclohex-1,4-ylene)NH(phen-1,4-ylene)(CH₂)₂—;

[0222] —(CH₂)₂C(O)NH(2-fluorophen-1,4-ylene)CH₂—;

[0223] —(CH₂)₂(phen-1,3-ylene)NH(phen-1,4-ylene)(CH₂)₂—;

[0224] —(CH₂)₂C(O)NH(2,5-difluorophen-1,4-ylene)CH₂—;

[0225] —(CH₂)₂NHC(O)(phen-1,4-ylene)(CH₂)₂—;

[0226] 1-[—CH₂(pyrid-2,6-ylene)CH₂](piperidin-4-yl)CH₂—;

[0227] —(CH₂)₃NH(phen-1,4-ylene)(CH₂)₂—;

[0228] —(CH₂)₂NH(naphth-1,4-ylene)(CH₂)₂—;

[0229] —(CH₂)₃O(phen-1,4-ylene)CH₂—;

[0230] 1-[—(CH₂)₃](piperidin-4-yl)CH₂—;

[0231] 4-[—(CH₂)₂](piperidin-1-yl)C(O)(phen-1,4-ylene)CH₂—;

[0232] —(CH₂)₃(phen-1,4-ylene)NHC(O)(CH₂)₂—;

[0233] —(CH₂)₃O(phen-1,4-ylene)(CH₂)₂—;

[0234] 2-[—(CH₂)₂](benzimidazol-5-yl)CH₂—;

[0235] —(CH₂)₂-(trans-cyclohex-1,4-ylene)NHC(O)(CH₂)₂—;

[0236] —(CH₂)₂-(trans-cyclohex-1,4-ylene)NHC(O)(CH₂)₄—;

[0237] —(CH₂)₂-(trans-cyclohex-1,4-ylene)NHC(O)(CH₂)₅—;

[0238] 4-[—(CH₂)₂](piperidin-1-yl)C(O)(CH₂)₂—;

[0239] —(CH₂)₂NHC(O)NH(phen-1,4-ylene)CH₂—;

[0240] —(CH₂)₂N(CH₃)(CH₂)₂(cis-cyclohex-1,4-ylene)-;

[0241] —(CH₂)₂C(O)NH(2,3,5,6-tetrafluorophen-1,4-ylene)CH₂—;

[0242] —(CH₂)₂C(O)NH(2,6-diiodophen-1,4-ylene)CH₂—;

[0243] 4-[—(CH₂)₂](piperidin-1-yl)C(O)(CH₂)₃—;

[0244] 4-[—(CH₂)₂](piperidin-1-yl)C(O)(CH₂)₄—;

[0245] 4-[—(CH₂)₂](piperidin-1-yl)C(O)(CH₂)₅—;

[0246] —(CH₂)₂C(O)NHCH₂(phen-1,4-ylene)CH₂—;

[0247] —(CH₂)₂NHC(O)NHCH₂(phen-1,4-ylene)CH₂—;

[0248] —(CH₂)₂C(O)NH(2-methylphen-1,4-ylene)CH₂—;

[0249] 1-[—(CH₂)₃0(phen-1,4-ylene)(CH₂)₂](piperidin-4-yl)CH₂—;

[0250] —(CH₂)₂C(O)NHCH₂(phen-1,3-ylene)(CH₂)₂—;

[0251] —(CH₂)₂O(phen-1,3-ylene)CH₂—;

[0252] —(CH₂)₂N(CH₃)C(O)CH₂O(phen-1,4-ylene)CH₂—;

[0253] —(CH₂)₂N(CH₃)C(O)CH₂O(phen-1,3-ylene)CH₂—;

[0254] —(CH₂)₂N(CH₃)C(O)(fur-2,5-ylene)CH₂—;

[0255] —(CH₂)₂N(CH₃)C(O)(thien-2,5-ylene)CH₂—;

[0256] —(CH₂)₂O(phen-1,4-ylene)O(CH₂)₂—;

[0257] —(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)(phen-1,4-ylene)CH₂—;

[0258] —(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)CH₂O(phen-1,2-ylene)CH₂—;

[0259] —(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)CH₂O(phen-1,3-ylene)CH₂—;

[0260] —(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)CH₂O(phen-1,4-ylene)CH₂—;

[0261] —(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)(fur-2,5-ylene)CH₂—;

[0262] —(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)(thien-2,5-ylene)CH₂—;

[0263] 4-[—(CH₂)₂](piperidin-1-yl)C(O)CH₂O(phen-1,2-ylene)CH₂—;

[0264] 4-[—(CH₂)₂](piperidin-1-yl)C(O)CH₂O(phen-1,3-ylene)CH₂—;

[0265] 4-[—(CH₂)₂](piperidin-1-yl)C(O)CH₂O(phen-1,4-ylene)CH₂—;

[0266] 4-[—(CH₂)₂](piperidin-1-yl)C(O)(fur-2,5-ylene)CH₂—;

[0267] 4-[—(CH₂)₂](piperidin-1-yl)C(O)(thien-2,5-ylene)CH₂—;

[0268] —(CH₂)₂(phen-1,4-ylene)NHC(O)(phen-1,3-ylene)CH₂—;

[0269] —(CH₂)₂(phen-1,4-ylene)NHC(O)(phen-1,4-ylene)CH₂—;

[0270] —(CH₂)₂(phen-1,4-ylene)NHC(O)CH₂O(phen-1,2-ylene)CH₂—;

[0271] —(CH₂)₂(phen-1,4-ylene)NHC(O)CH₂O(phen-1,3-ylene)CH₂—;

[0272] —(CH₂)₂(phen-1,4-ylene)NHC(O)CH₂O(phen-1,4-ylene)CH₂—;

[0273] —(CH₂)₂(phen-1,4-ylene)NHC(O)(fur-2,5-ylene)CH₂—;

[0274] —(CH₂)₂(phen-1,4-ylene)NHC(O)(thien-2,5-ylene)CH₂—;

[0275] —(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)(phen-1,3-ylene)CH₂—;

[0276] —(CH₂)₃O(phen-1,3-ylene)CH₂—;

[0277] —CH₂CH(OH)CH₂NH(phen-1,4-ylene)(CH₂)₂—;

[0278] —(CH₂)₄NH(phen-1,4-ylene)(CH₂)₂—;

[0279] —(CH₂)₂C(O)NH(phen-1,4-ylene)CH₂NHC(O)CH₂—;

[0280] —(CH₂)₂C(O)NH(phen-1,4-ylene)(CH₂)₂NHC(O)CH₂—;

[0281] —(CH₂)₂C(O)NHCH₂(trans-cyclohex-1,4-ylene)CH₂—;

[0282] —(CH₂)₂NHC(O)(CH₂)₅—;

[0283] —(CH₂)₂O(phen-1,3-ylene)O(CH₂)₂—;

[0284] —(CH₂)₂O(phen-1,2-ylene)O(CH₂)₂—;

[0285] —CH₂(phen-1,2-ylene)O(phen-1,2-ylene)CH₂—;

[0286] —(CH₂)₂C(O)NH(CH₂)₆—;

[0287] —(CH₂)₃(phen-1,4-ylene)(CH₂)₃—;

[0288] —(CH₂)₃(phen-1,4-ylene)(CH₂)₂—;

[0289] —(CH₂)₄(phen-1,4-ylene)(CH₂)₂—;

[0290] —(CH₂)₃(furan-2,5-ylene)(CH₂)₃—;

[0291] —(CH₂)₂N(CH₃)C(O)NH(phen-1,4-ylene)(CH₂)₂—;

[0292] 4-[—(CH₂)₂](piperidin-1-yl)C(O)NH(phen-1,4-ylene)(CH₂)₂—;

[0293] —(CH₂)₃(phen-1,3-ylene)(CH₂)₃—;

[0294] —(CH₂)₃(tetrahydrofuran-2,5-ylene)(CH₂)₃—; and

[0295] —(CH₂)₂O(phen-1,4-ylene)C(O)(CH₂)₂—.

[0296] Representative Subgeneric Groupings

[0297] The following subgeneric formulae and groupings are intended toprovide representative examples of various aspects and embodiments ofthis invention and as such, they are not intended to exclude otherembodiments or to limit the scope of this invention unless otherwiseindicated. In particular, these subgeneric formulae and groupingsprovide representative examples of compounds of formula I that may befound in particular libraries of this invention.

[0298] A particular group of compounds of formula I are those disclosedin U.S. Provisional Application No. 60/447,843, filed on Feb. 14, 2003.This group includes compounds of formula I; wherein:

[0299] a is 0 or an integer of from 1 to 3;

[0300] each R¹ is independently selected from (1-4C)alkyl,(2-4C)alkenyl, (2-4C)alkynyl, (3-6C)cycloalkyl, cyano, halo, —OR^(1a),—C(O)OR^(1b), SR^(1c), —S(O)R^(1d), —S(O)₂R^(1e) and —NR^(1f)R^(1g);

[0301] each of R^(1a), R^(1b), R^(1c), R^(1d), R^(1e), R^(1f) and R^(1g)is independently hydrogen or (1-4C)alkyl;

[0302] b is o or an integer of from 1 to 3;

[0303] each R² is independently selected from (1-4C)alkyl,(2-4C)alkenyl, (2-4C)alkynyl, (3-6C)cycloalkyl, cyano, halo, —OR^(2a),—C(O)OR^(2b), SR^(2c), —S(O)R^(2d), S(O)₂R^(2e) and —NR^(2f)R^(2g);

[0304] each of R^(2a), R^(2b), R^(2c), R^(2d), R^(2e), R^(2f) and R^(2g)is independently hydrogen or (1-4C)alkyl;

[0305] W is attached to the 3- or 4-position with respect to thenitrogen atom in the piperidine ring, and represents O or NW^(a);

[0306] W^(a) is hydrogen or (1-4C)alkyl;

[0307] c is O or an integer of from 1 to 4;

[0308] each R³ is a substituent on carbon independently selected from(1-4C)alkyl, (2-4C)alkenyl, (2-4C)alkynyl, (3-6C)cycloalkyl, cyano,halo, —OR^(3a), —C(O)OR^(3b), SR^(3c), —S(O)R^(3d), —S(O)₂R^(3e) and—NR^(3f)R^(3g);

[0309] each of R^(3a), R^(3b), R^(3c), R^(3d), R^(3e), R^(3f) and R^(3g)is independently hydrogen or (1-4C)alkyl;

[0310] R⁴ is a divalent group of the formula:

—(R^(4a))_(d)—(A¹)_(e)—(R^(4b))_(f)—Q—(R^(4c))_(g)—(A²)_(h)—(R^(4d))_(i)—

[0311] wherein

[0312] d, e, f, g, h and i are each independently selected from 0 and 1;

[0313] R^(4a), R^(4b), R^(4c) and R^(4d) are each independently selectedfrom (1-10C)alkylene, (2-10C)alkenylene and (2-10C)alkynylene whereineach alkylene, alkenylene or alkynylene group is unsubstituted orsubstituted with from 1 to 5 substituents independently selected from(1-4C)alkyl, fluoro, hydroxy, phenyl and phenyl(l-4C)-alkyl;

[0314] A¹ and A² are each independently selected from(3-7C)cycloalkylene, (6-10C)arylene, (2-9C)heteroarylene and(3-6C)heterocyclene; wherein each cycloalkylene is unsubstituted orsubstituted with from 1 to 4 substitutents selected independently from(1-4C)alkyl and each arylene, heteroarylene or heterocyclene group isunsubstituted or substituted with from 1 to 4 substituents independentlyselected from halo, (1-4C)alkyl and (1-4C)alkoxy;

[0315] Q is selected from a bond, —O—, —C(O)O—, —OC(O)—, —S—, —S(O)—,—S(O)₂—, —N(Q^(a))C(O)—, —C(O)N(Q^(b))—, —N(Q^(c))S(O)₂—,—S(O)₂N(Q^(d))—, —N(Q^(e))C(O)N(Q^(f))—, —N(Q^(g))S(O)₂N(Q^(h))—,—OC(O)N(Q^(i))— and —N(Q^(j))C(O)O—;

[0316] Q^(a), Q^(b), Q^(c), Q^(d), Q^(e), Q^(f), Q^(g), Q^(h), Q^(i) andQ^(j) are each independently selected from hydrogen, (1-6C)alkyl, A³ and(1-4C)alkylene-A⁴; wherein the alkyl group is unsubstituted orsubstituted with from 1 to 3 substituents independently selected fromfluoro, hydroxy and (1-4C)alkoxy; or together with the nitrogen atom andthe group R^(4b) or R^(4c) to which they are attached, form a 4-6membered azacycloalkylene group;

[0317] A³ and A⁴ are each independently selected from (3-6C)cycloalkyl,(6-10C)aryl, (2-9C)heteroaryl and (3-6C)heterocyclyl; wherein eachcycloalkyl is unsubstituted or substituted with from 1 to 4substitutents selected independently from (1-4C)alkyl and each aryl,heteroaryl or heterocyclyl group is unsubstituted or substituted withfrom 1 to 4 substituents independently selected from halo, (1-4C)alkyland (1-4C)alkoxy;

[0318] provided that the number of contiguous atoms in the shortestchain between the two nitrogen atoms to which R⁴ is attached is in therange of from 8 to 14;

[0319] R⁵ represents hydrogen or (1-4C)alkyl;

[0320] R⁶ is —NR^(6a)CR^(6b)(O) and R⁷ is hydrogen, or R⁶ and R⁷together form —NR^(7a)C(O)—CR^(7b)═CR^(7c)—,—CR^(7d)═CR^(7e)—C(O)—NR^(7f)—,—NR^(7g)C(O)—CR^(7h)R^(7i)—CR^(7j)R^(7k)— or—CR^(7l)R^(7m)—CR^(7n)R^(7o)—C(O)—NR^(7p)—;

[0321] each of R^(6a) and R^(6b) is independently hydrogen or(1-4C)alkyl; and

[0322] each of R^(7a), R^(7b), R^(7c), R_(7d), R^(7e), R^(7f), R^(7g),R^(7h), R^(7i), R^(7j), R^(7k), R^(7l), R^(7m), R^(7n), R^(7o), andR^(7p) is independently hydrogen or (1-4C)alkyl;

[0323] or a pharmaceutically acceptable salt or solvate or stereoisomerthereof.

[0324] Another particular group of compounds of formula I are thosedisclosed in U.S. Provisional Application No. 60/467,035, filed on May1, 2003. This group of compounds includes compounds of formula I;wherein:

[0325] a is 0 or an integer of from 1 to 3;

[0326] each R¹ is independently selected from (1-4C)alkyl,(2-4C)alkenyl, (2-4C)alkynyl, (3-6C)cycloalkyl, cyano, halo, —OR^(1a),—C(O)OR^(1b), SR^(1c), —S(O)R^(1d), —S(O)₂R^(1e), and —NR^(1f)R^(1g);

[0327] each of R^(1a), R^(1b), R^(1c), R^(1d), R^(1e), R^(1f) and R^(1g)is independently hydrogen or (1-4C)alkyl;

[0328] b is 0 or an integer of from 1 to 3;

[0329] each R² is independently selected from (1-4C)alkyl,(2-4C)alkenyl, (2-4C)alkynyl, (3-6C)cycloalkyl, cyano, halo, —OR^(2a),—C(O)OR^(2b), SR^(2c), S(O)R^(2d), S(O)₂R^(2e), and —NR^(2f)R^(2g);

[0330] each of R^(2a), R^(2b), R^(2c), R^(2d), R^(2e), R^(2f) and R^(2g)is independently hydrogen or (1-4C)alkyl;

[0331] W is attached to the 3- or 4-position with respect to thenitrogen atom in the piperidine ring, and represents O or NW^(a);

[0332] W^(a) is hydrogen or (1-4C)alkyl;

[0333] c is 0 or an integer of from 1 to 4;

[0334] each R³ is a substituent on carbon independently selected from(1-4C)alkyl, (2-4C)alkenyl, (2-4C)alkynyl, (3-6C)cycloalkyl, cyano,halo, —OR^(3a), —C(O)OR^(3b), SR^(3c), —S(O)R^(3d), —S(O)₂R^(3e), and—NR^(3f)R^(3g);

[0335] each of R^(3a), R^(3b), R^(3c), R^(3d), R^(3e), R^(3f) and R^(3g)is independently hydrogen or (1-4C)alkyl;

[0336] R⁴ is a divalent group of the formula:

—(R^(4a))_(d)—(A¹)_(e)—(R^(4b))_(f)—Q—(R^(4c))_(g)—(A²)_(h)—(R^(4d))_(i)—

[0337] wherein

[0338] d, e, f, g, h and i are each independently selected from 0 and 1;

[0339] R^(4a), R^(4b), R^(4c) and R^(4d) are each independently selectedfrom (1-10C)alkylene, (2-10C)alkenylene and (2-10C)alkynylene whereineach alkylene, alkenylene or alkynylene group is unsubstituted orsubstituted with from 1 to 5 substituents independently selected from(1-4C)alkyl, fluoro, hydroxy, phenyl and phenyl(1-4C)-alkyl;

[0340] A¹ and A² are each independently selected from(3-7C)cycloalkylene, (6-10C)arylene, (2-9C)heteroarylene and(3-6C)heterocyclene; wherein each cycloalkylene is unsubstituted orsubstituted with from 1 to 4 substitutents selected independently from(1-4C)alkyl and each arylene, heteroarylene or heterocyclene group isunsubstituted or substituted with from 1 to 4 substituents independentlyselected from halo, (1-4C)alkyl and (1-4C)alkoxy;

[0341] Q is selected from a bond, —O—, —C(O)O—, —OC(O)—, —S—, —S(O)—,—S(O)₂—, —N(Q^(a))C(O)—, —C(O)N(Q^(b))—, —N(Q^(c))S(O)₂—,—S(O)₂N(Q^(d))—, —N(Q^(e))C(O)N(Q^(f))—, —N(Q^(g))S(O)₂N(Q^(h))—,—OC(O)N(Q^(i))— and —N(Q^(j))C(O)O—;

[0342] Q^(a), Q^(b), Q^(c), Q^(d), Q^(e), Q^(f), Q^(g), Q^(h), Q^(i) andQ^(j) are each independently selected from hydrogen, (1-6C)alkyl, A³ and(1-4C)alkylene-A⁴; wherein the alkyl group is unsubstituted orsubstituted with from 1 to 3 substituents independently selected fromfluoro, hydroxy and (1-4C)alkoxy; or together with the nitrogen atom andthe group R^(4b) or R^(4c) to which they are attached, form a 4-6membered azacycloalkylene group;

[0343] A³ and A⁴ are each independently selected from (3-6C)cycloalkyl,(6-10C)aryl, (2-9C)heteroaryl and (3-6C)heterocyclyl; wherein eachcycloalkyl is unsubstituted or substituted with from 1 to 4substitutents selected independently from (1-4C)alkyl and each aryl,heteroaryl or heterocyclyl group is unsubstituted or substituted withfrom 1 to 4 substituents independently selected from halo, (1-4C)alkyland (1-4C)alkoxy;

[0344] provided that the number of contiguous atoms in the shortestchain between the two nitrogen atoms to which R⁴ is attached is in therange of from 4 to 14;

[0345] R⁵ represents hydrogen or (1-4C)alkyl;

[0346] R⁶ is —NR^(6a)CR^(6b)(O) or CR^(6c)R^(6d)OR^(6e) and R⁷ ishydrogen, or R⁶ and R⁷ together form —NR^(7a)C(o)—CR^(7b)═CR^(7c)—,—CR^(7d)═CR^(7e)—C(O)—NR^(7f)—,—NR^(7g)C(O)—CR^(7h)R^(7i)—CR^(7j)R^(7k)— or—CR^(7l)R^(7m)—CR^(7n)R^(7o)—C(O)—NR^(7p)—;

[0347] each of R^(6a), R^(6b), R^(6c), R^(6d) and R^(6e) isindependently hydrogen or (1-4C)alkyl; and

[0348] each of R^(7a), R^(7b), R^(7c), R^(7d), R^(7e), R^(7f), R^(7g),R^(7h), R^(7i), R^(7j), R^(7k), R^(7l), R^(7m), R^(7n), R^(7o), andR^(7p) is independently hydrogen or (1-4C)alkyl;

[0349] or a pharmaceutically acceptable salt or solvate or stereoisomerthereof.

[0350] Another particular group of compounds of formula I are thosewhere: a is 0; b is 0; c is 0; W is O; W is attached at the 4-positionof the piperidinyl ring; R⁵ is hydrogen; and R⁴, R⁶ and R⁷ are asdefined herein; or a salt or stereoisomer thereof.

[0351] Still another particular group of compounds of formula I arethose wherein: a is 0; b is 0; c is 0; W is NH; W is attached at the4-position of the piperidinyl ring; R⁵ is hydrogen; and R⁴, R⁶ and R⁷are as defined herein; or a salt or stereoisomer thereof.

[0352] Yet another particular group of compounds of formula I are thosewherein: a is 0; b is 0; c is 0; W is O; W is attached at the 4-positionof the piperidinyl ring; R⁴ is —(CH₂)_(j)— where j is 8, 9 or 10; R⁵ ishydrogen; and R⁶ and R⁷ are as defined herein; or a salt or stereoisomerthereof.

[0353] Another particular group of compounds of formula I are thosewherein: a is 0; b is 0; c is 0; W is NH;.W is attached at the4-position of the piperidinyl ring; R⁴ is —(CH₂)_(j)— where j is 8, 9 or10; R⁵ is hydrogen; and R⁶ and R⁷ are as defined herein; or a salt orstereoisomer thereof.

[0354] Yet another particular group of compounds of formula I are thosewherein: a is 0; b is 0; c is 0; W is O; W is attached at the 4-positionof the piperidinyl ring; R⁴ is —(CH₂)₂— C(O)NH—(CH₂)₅—; R⁵ is hydrogen;and R⁶ and R⁷ are as defined herein; or a salt or stereoisomer thereof.

[0355] Another particular group of compounds of formula I are thosewherein: a is 0; b is 0; c is 0; W is NH; W is attached at the4-position of the piperidinyl ring; R⁴ is —(CH₂)₂— C(O)NH—(CH₂)₅—; R⁵ ishydrogen; and R⁶ and R⁷ are as defined herein; or a salt or stereoisomerthereof.

[0356] Another particular group of compounds of formula I are those offormula II as defined herein; or a salt or stereoisomer thereof.

[0357] Another particular group of compounds of formula I are those offormula III as defined herein; or a salt or stereoisomer thereof.

[0358] Another particular group of compounds of formula I are those offormula IV as defined herein; or a salt or stereoisomer thereof.

[0359] Another particular group of compounds of formula I are those offormula II, III or IV as defined herein, wherein the piperidinyl ring issubstituted at the 4-position with a methyl group; or a salt orstereoisomer thereof.

[0360] Representative compounds of formula I include compounds offormula V:

[0361] wherein W, R⁴, R⁶ and R⁷ are as defined in Table I; or apharmaceutically acceptable salt or solvate thereof. TABLE I Ex. W R⁴ R⁶R⁷ 1 NH —(CH₂)₉— (racemic)¹ —NHC(O)CH═CH—² 2 O —(CH₂)₉— (racemic)—NHC(O)CH═CH— 3 O —(CH₂)₉— —NHC(O)CH═CH— 4 O —(CH₂)₉— H —NHC(O)H 5 O—(CH₂)₉— —NHC(O)CH₂CH₂— 6 O —(CH₂)₂C(O)NH(CH₂)₅— —NHC(O)CH═CH— 7 O—(CH₂)₂N(CH₃)C(O)(CH₂)₅— —NHC(O)CH═CH— 8 O—(CH₂)₂C(O)NH(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 9 O—(CH₂)₂NHC(O)(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 10 O—(CH₂)₂NHC(O)NH(CH₂)₅— —NHC(O)CH═CH— 11 O —(CH₂)₃NHC(O)NH(CH₂)₅——NHC(O)CH═CH— 12 O —(CH₂)₉— H —CH₂OH 13 NH —(CH₂)₉— H —CH₂OH 14 O—(CH₂)₂C(O)NHCH₂(cyclohex-1,3-ylene)CH₂— —NHC(O)CH═CH— 15 O—(CH₂)₂NHC(O)(cis-cyclopent-1,3-ylene)— —NHC(O)CH₂CH₂— 16 O—(CH₂)₂C(O)NH(2-chlorophen-1,4-ylene)CH₂— —NHC(O)CH═CH— 17 O—(CH₂)₂S(O)₂NH(CH₂)₅— —NHC(O)CH═CH— 18 O—(CH₂)₂N(CH₃)S(O)₂(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 19 O—(CH₂)₂NHC(O)NHCH₂(phen-1,3-ylene)CH₂— —NHC(O)CH═CH— 20 O—(CH₂)₃(phen-1,4-ylene)NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 21 O1-[—CH₂(2-fluorophen-1,3-ylene)CH₂](piperidin-4-yl)CH₂— —NHC(O)CH═CH— 22O —(CH₂)₃O(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 23 O—(CH₂)₂(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 24 O—(CH₂)₃(thien-2,5-ylene)(CH₂)₃— —NHC(O)CH═CH— 25 O—(CH₂)₂C(O)NH(2-chloro-5-methoxyphen-1,4-ylene)CH₂— —NHC(O)CH═CH— 26 O—(CH₂)₇— —NHC(O)CH═CH— 27 O —(CH₂)₈— —NHC(O)CH═CH— 28 O—(CH₂)₂NHC(O)NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 29 O1-[—(CH₂)₂C(O)](piperidin-4-yl)(CH₂)₂— —NHC(O)CH═CH— 30 O—(CH₂)₂NHC(O)(trans-cyclohex-1,4-ylene)CH₂— —NHC(O)CH═CH— 31 O—(CH₂)₂NHC(O)(cis-cyclopent-1,3-ylene)- —NHC(O)CH═CH— 32 O—(CH₂)₂NHC(O)NH(CH₂)₅— H —NHC(O)H 33 O —(CH₂)₂NH(phen-1,4-ylene)(CH₂)₂——NHC(O)CH═CH— 34 O —(CH₂)₃NHC(O)NH(CH₂)₅— H —NHC(O)H 35 O1-[—(CH₂)₂NHC(O)](piperidin-4-yl)(CH₂)₂— —NHC(O)CH═CH— 36 O—CH₂(phen-1,4-ylene)NH(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 37 O—(CH₂)₂C(O)NHCH₂(phen-1,3-ylene)CH₂— —NHC(O)CH═CH— 38 NH—(CH₂)₂C(O)NH(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 39 O—(CH₂)₂C(O)NHCH₂(pyrid-2,6-ylene)CH₂— —NHC(O)CH═CH— 40 O—(CH₂)₂C(O)NH(cis-cyclohex-1,4-ylene)CH₂— —NHC(O)CH═CH— 41 O—(CH₂)₂C(O)NH(trans-cyclohex-1,4-ylene)CH₂— —NHC(O)CH═CH— 42 O—(CH₂)₂NHC(O)(cis-cyclopent-1,3-ylene)CH₂— —NHC(O)CH═CH— 43 O—(CH₂)₂N(CH₃)C(O)(phen-1,3-ylene)CH₂— —NHC(O)CH═CH— 44 O—(CH₂)₂N(CH₃)C(O)(trans-cyclohex-1,4-ylene)CH₂— —NHC(O)CH—CH— 45 O—(CH₂)₂C(O)NH(phen-1,4-ylene)CH₂— (racemic) —NHC(O)CH═CH— 46 O—(CH₂)₂C(O)NH(phen-1,4-ylene)C*H(CH₃)—((S)-isomer) —NHC(O)CH═CH— 47 O—(CH₂)₂C(O)NH(phen-1,4-ylene)C*H(CH₃)-((R)-isomer) —NHC(O)CH═CH— 48 O2-[(S)-(—CH₂-](pyrrolidin-1-yl)C(O)(CH₂)₄— —NHC(O)CH═CH— 49 O2-[(S)-(—CH₂-](pyrrolidin-1-yl)C(O)(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 50O —(CH₂)₂C(O)NH(phen-1,4-ylene)CH₂— H —NHC(O)H 51 O—(CH₂)₂C(O)NH(phen-1,4-ylene)C*H(CH₃)-((R)-isomer) H —NHC(O)H 52 O—(CH₂)₂C(O)NH(4-chlorophen-1,3-ylene)CH₂— —NHC(O)CH═CH— 53 NH—(CH₂)₂C(O)NH(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 54 NH1-[—(CH₂)₂C(0)](piperidin-4-yl)(CH₂)₂— —NHC(O)CH═CH— 55 NH—(CH₂)₂C(O)NHCH₂(phen-1,3-ylene)CH₂— —NHC(O)CH═CH— 56 O—CH₂(2-fluorophen-1,3-ylene)CH₂— —NHC(O)CH═CH— 57 O—(CH₂)₂C(O)NH(4-methylphen-1,3-ylene)CH₂— —NHC(O)CH═CH— 58 O—(CH₂)₂C(O)NH(6-chlorophen-1,3-ylene)CH₂— —NHC(O)CH═CH— 59 O—(CH₂)₂C(O)NH(2,6-dichlorophen-1,4-ylene)CH₂— —NHC(O)CH═CH— 60 O4-[—CH₂-](piperidin-1-yl)C(O)(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 61 O—(CH₂)₂NHC(O)(phen-1,4-ylene)CH₂— H —NHC(O)H 62 O—(CH₂)₂C(O)N(CH₂CH₃)(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 63 O1-[—(CH₂)₂NHC(O)](piperidin-4-yl)- —NHC(O)CH═CH— 64 O—(CH₂)₂C(O)NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 65 O—(CH₂)₂NHC(O)(thien-2,5-ylene)CH₂— —NHC(O)CH═CH— 66 O—(CH₂)2N(CH₃)C(O)(3-nitrophen-1,4-ylene)CH₂— —NHC(O)CH═CH— 67 O—(CH₂)₂C(O)NH(trans-cyclohex-1,4-ylene)CH₂— H —NHC(O)H 68 O—(CH₂)₂N(CH₃)C(O)(trans-cyclohex-1,4-ylene)— —NHC(O)CH═CH— 69 O1-[—CH₂(2-fluorophen-1,3-ylene)CH₂](piperidin-4-yl)- —NHC(O)CH═CH— 70 O5-[—(CH₂)₂NHC(O)](pyrid-2-yl)CH₂— —NHC(O)CH═CH— 71 O1-[—(CH₂)₃](piperidin-4-yl)CH₂— —NHC(O)CH═CH— 72 O—(CH₂)₂C(O)NH(phen-1,4-ylene)(CH₂)₂— H —NHC(O)H 73 O—CH₂(phen-1,2-ylene)NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 74 O1-[—CH₂(2-fluorophen-1,3-ylene)CH₂](piperidin-4-yl)(CH₂)₂— —NHC(O)CH═CH—75 O —(CH₂)₃NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 76 O—(CH₂)₂C(O)NH(3-chlorophen-1,4-ylene)CH₂— —NHC(O)CH═CH— 77 O—(CH₂)₂C(O)NH(2-(CF₃O-)phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 78 O—(CH₂)₃(phen-1,3-ylene)NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 79 O—CH₂(phen-1,3-ylene)NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 80 O—(CH₂)₂C(O)NH(2-iodophen-1,4-ylene)CH₂— —NHC(O)CH═CH— 81 O—(CH₂)₂C(O)NH(2-chloro-6-methylphen-1,4-ylene)CH₂— —NHC(O)CH═CH— 82 O—(CH₂)₂C(O)NH(CH₂)₅— (racemic) —NHC(O)CH═CH— 83 O—(CH₂)₂C(O)NH(2-bromophen-1,4-ylene)CH₂— —NHC(O)CH═CH— 84 O—(CH₂)₃(phen-1,2-ylene)NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 85 O1-[—CH₂(2-fluorophen-1,3-ylene)CH₂](piperidin-4-yl)(CH₂)₃— —NHC(O)CH═CH—86 O —(CH₂)₂C(O)NH(2-methoxyphen-1,4-ylene)CH₂— —NHC(O)CH═CH— 87 O—(CH₂)₅NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 88 O4-[—(CH₂)₂-](piperidin-1-yl)(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 89 O—(CH₂)₂C(O)NH(phen-1,4-ylene)CH(CH₃)CH₂— —NHC(O)CH═CH— 90 O—(CH₂)₂-(trans-cyclohex-1,4-ylene)NH(phen-1,4-ylene)(CH₂)₂——NHC(O)CH═CH— 91 O —(CH₂)₂C(O)NH(2-fluorophen-1,4-ylene)CH₂——NHC(O)CH═CH— 92 O —(CH₂)₂(phen-1,3-ylene)NH(phen-1,4-ylene)(CH₂)₂——NHC(O)CH═CH— 93 O —(CH₂)₂C(O)NH(2,5-difluorophen-1,4-ylene)CH₂——NHC(0)CH═CH— 94 O —(CH₂)₂NHC(O)(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 95O 1-[—CH₂(pyrid-2,6-ylene)CH₂](piperidin-4-yl)CH₂— —NHC(O)CH═CH— 96 O—(CH₂)₂NH(naphth-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 97 O4-[—(CH₂)₂](piperidin-1-yl)C(O)(phen-1,4-ylene)CH₂— —NHC(0)CH═CH— 98 O—(CH₂)₃(phen-1,4-ylene)NHC(O)(CH₂)₂— —NHC(O)CH═CH— 99 O—(CH₂)₃O(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 100 O2-[—(CH₂)₂](benzimidazol-5-yl)CH₂— —NHC(O)CH═CH— 101 O—(CH₂)₂-(trans-cyclohex-1,4-ylene)NHC(O)(CH₂)₂— —NHC(O)CH═CH— 102 O—(CH₂)₂-(trans-cyclohex-1,4-ylene)NHC(O)(CH₂)₄— —NHC(O)CH═CH— 103 O—(CH₂)₂-(trans-cyclohex-1,4-ylene)NHC(O)(CH₂)₅— —NHC(O)CH═CH— 104 O4-[—(CH₂)₂](piperidin-1-yl)C(O)(CH₂)₂— —NHC(O)CH═CH— 105 O—(CH₂)₂NHC(O)NH(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 106 O—(CH₂)₂N(CH₃)(CH₂)₂(cis-cyclohex-1,4-ylene)- —NHC(O)CH═CH— 107 O—(CH₂)₂C(O)NH(2,3,5,6-tetrafluorophen-1,4-ylene)CH₂— —NHC(O)CH═CH— 108 O—(CH₂)₂C(O)NH(2,6-diiodophen-1,4-ylene)CH₂— —NHC(O)CH═CH— 109 O4-[—(CH₂)₂](piperidin-1-yl)C(O)(CH₂)₃— —NHC(O)CH═CH— 110 O4-[—(CH₂)₂](piperidin-1-yl)C(O)(CH₂)₄— —NHC(O)CH═CH— 111 O4-[—(CH₂)₂](piperidin-1-yl)C(O)(CH₂)₅— —NHC(O)CH═CH— 112 O—(CH₂)₂C(O)NHCH₂(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 113 O—(CH₂)₂C(O)NHCH₂(phen-1,4-ylene)CH₂— H —NHC(O)H 114 O—(CH₂)₂NHC(O)NHCH₂(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 115 O—(CH₂)₂NHC(O)NHCH₂(phen-1,4-ylene)CH₂— H —NHC(O)H 116 O—(CH₂)₂C(O)NH(2-methylphen-1,4-ylene)CH₂— —NHC(O)CH═CH— 117 O1-[—(CH₂)₃O(phen-1,4-ylene)(CH₂)₂](piperidin-4-yl)CH₂— —NHC(O)CH═CH— 118O —(CH₂)₂C(O)NHCH₂(phen-1,3-ylene)(CH₂)₂— —NHC(O)CH═CH— 119 O—(CH₂)₂O(phen-1,3-ylene)CH₂— —NHC(O)CH═CH— 120 O—(CH₂)₂N(CH₃)C(O)CH₂O(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 121 O—(CH₂)₂N(CH₃)C(O)CH₂O(phen-1,3-ylene)CH₂— —NHC(O)CH═CH— 122 O—(CH₂)₂N(CH₃)C(O)(fur-2,5-ylene)CH₂— —NHC(O)CH═CH— 123 O—(CH₂)₂N(CH₃)C(O)(thien-2,5-ylene)CH₂— —NHC(O)CH═CH— 124 O—(CH₂)₂O(phen-1,4-ylene)O(CH₂)₂— —NHC(O)CH═CH— 125 O—(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)(phen-1,4-ylene)CH₂——NHC(O)CH═CH— 126 O—(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)CH₂O(phen-1,2-ylene)CH₂——NHC(O)CH═CH— 127 O—(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)CH₂O(phen-1,3-ylene)CH₂——NHC(O)CH═CH— 128 O—(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)CH₂O(phen-1,4-ylene)CH₂——NHC(O)CH═CH— 129 O—(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)(fur-2,5-ylene)CH₂— —NHC(O)CH═CH—130 O —(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)(thien-2,5-ylene)CH₂——NHC(O)CH═CH— 131 O4-[—(CH₂)₂](piperidin-1-yl)C(O)CH₂O(phen-1,2-ylene)CH₂— —NHC(O)CH═CH—132 O 4-[—(CH₂)₂](piperidin-1-yl)C(O)CH₂O(phen-1,3-ylene)CH₂——NHC(O)CH═CH— 133 O4-[—(CH₂)₂](piperidin-1-yl)C(O)CH₂O(phen-1,4-ylene)CH₂— —NHC(O)CH═CH—134 O 4-[—(CH₂)₂](piperidin-1-yl)C(O)(fur-2,5-ylene)CH₂— —NHC(O)CH═CH—135 O 4-[—(CH₂)₂](piperidin-1-yl)C(O)(thien-2,5-ylene)CH₂— —NHC(O)CH═CH—136 O —(CH₂)₂(phen-1,4-ylene)NHC(O)(phen-1,3-ylene)CH₂— —NHC(O)CH═CH—137 O —(CH₂)₂(phen-1,4-ylene)NHC(O)(phen-1,4-ylene)CH₂— —NHC(O)CH═CH—138 O —(CH₂)₂(phen-1,4-ylene)NHC(O)CH₂O(phen-1,2-ylene)CH₂——NHC(O)CH═CH— 139 O—(CH₂)₂(phen-1,4-ylene)NHC(O)CH₂O(phen-1,3-ylene)CH₂— —NHC(O)CH═CH— 140O —(CH₂)₂(phen-1,4-ylene)NHC(O)CH₂O(phen-1,4-ylene)CH₂— —NHC(O)CH═CH—141 O —(CH₂)₂(phen-1,4-ylene)NHC(O)(fur-2,5-ylene)CH₂— —NHC(O)CH═CH— 142O —(CH₂)₂(phen-1,4-ylene)NHC(O)(thien-2,5-ylene)CH₂— —NHC(O)CH═CH— 143 O—(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)(phen-1,3-ylene)CH₂——NHC(O)CH═CH— 144 O —(CH₂)₃O(phen-1,3-ylene)CH₂— —NHC(O)CH═CH— 145 O—CH₂CH(OH)CH₂NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 146 O—(CH₂)₄NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 147 O—(CH₂)₂C(O)NH(phen-1,4-ylene)CH₂NHC(O)CH₂— —NHC(O)CH═CH— 148 O—(CH₂)₂C(O)NH(phen-1,4-ylene)(CH₂)₂NHC(O)CH₂— —NHC(O)CH═CH— 149 O—(CH₂)₂C(O)NHCH₂(trans-cyclohex-1,4-ylene)CH₂— —NHC(O)CH═CH— 150 O—(CH₂)₂NHC(O)(CH₂)₅— —NHC(O)CH═CH— 151 O—(CH₂)₂O(phen-1,3-ylene)O(CH₂)₂— —NHC(O)CH═CH— 152 O—(CH₂)₂O(phen-1,2-ylene)O(CH₂)₂— —NHC(O)CH═CH— 153 O—CH₂(phen-1,2-ylene)O(phen-1,2-ylene)CH₂— —NHC(O)CH═CH— 154 O—(CH₂)₂C(O)NH(CH₂)_(6—) —NHC(O)CH═CH— 155 O—(CH₂)₂NHC(O)(cis-cyclopent-1,3-ylene)- —NHC(O)CH═CH— 156 O—(CH₂)₃(phen-1,4-ylene)(CH₂)₃— —NHC(O)CH═CH— 157 O—(CH₂)₃(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 158 O—(CH₂)₄(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 159 O—(CH₂)₃(furan-2,5-ylene)(CH₂)₃— —NHC(O)CH═CH— 160 O—(CH₂)₂N(CH₃)C(O)NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 161 O4-[—(CH₂)₂](piperidin-1-yl)C(O)NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH—162 O —(CH₂)₃(phen-1,3-ylene)(CH₂)₃— —NHC(O)CH═CH— 163 O—(CH₂)₃(tetrahydrofuran-2,5-ylene)(CH₂)₃— —NHC(O)CH═CH— 164 O—(CH₂)₂O(phen-1,4-ylene)C(O)(CH₂)₂— —NHC(O)CH═CH—

[0362] Another particular group of compounds of formula I are compoundsof formula VI:

[0363] wherein W, R^(1A), R^(1B), R^(1C), R^(2A), R^(2B), R⁴, R⁶ and R⁷are as defined in Table II; or a pharmaceutically acceptable salt orsolvate thereof. TABLE II Ex. W R^(1A) R^(1B) R^(1C) R^(2A) R^(2B) R⁴ R⁶R⁷ 165 O H H H Br H —(CH₂)₉— (racemic) —NHC(O)CH═CH— 166 O F H H H H—(CH₂)₉— —NHC(O)CH═CH— 167 O H Cl H F F —(CH₂)₉— —NHC(O)CH═CH— 168 O HCl Cl F F —(CH₂)₉— —NHC(O)CH═CH— 169 O H H H F F —(CH₂)₉— —NHC(O)CH═CH—

[0364] Another particular group of compounds of formula I are compoundsof formula VII:

[0365] wherein W, R⁴, R⁶ and R⁷ are as defined in Table III; or apharmaceutically acceptable salt or solvate thereof. TABLE III Ex. W R⁴R⁶ R⁷ 170 O —(CH₂)₂C(O)NH(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 171 O—(CH₂)₂C(O)NH(phen-1,4-ylene)CH₂— H —NHC(O)H 172 O —(CH₂)₉——NHC(O)CH═CH— 173 O —(CH₂)₉— H —NHC(O)H 174 O —(CH₂)₂C(O)NH(CH₂)₅——NHC(O)CH₂CH₂— 175 O —(CH₂)₂C(O)NH(CH₂)₅— H —NHC(O)H 176 O—(CH₂)₂NHC(O)(CH₂)₅— —NHC(O)CH═CH— 177 O —(CH₂)₂NHC(O)(CH₂)₅— H —NHC(O)H178 O —(CH₂)₂NHC(O)(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 179 O—(CH₂)₂NHC(O)(phen-1,4-ylene)CH₂— H —NHC(O)H 180 O—(CH₂)₃(phen-1,4-ylene)NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 181 O—(CH₂)₂NHC(O)(2-chlorophen-1,4-ylene)CH₂— —NHC(O)CH═CH— 182 O—(CH₂)₂NHC(O)(2-chloro-5-methoxyphen-1,4-ylene)CH₂— —NHC(O)CH═CH—

[0366] Definitions

[0367] When describing the libraries, compounds, compositions, methodsand processes of this invention, the following terms have the followingmeanings unless otherwise indicated.

[0368] The term “alkyl” means a monovalent saturated hydrocarbon groupwhich may be linear or branched. Unless otherwise defined, such alkylgroups typically contain from 1 to 10 carbon atoms. Representative alkylgroups include, by way of example, methyl, ethyl, n-propyl, isopropyl,n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, n-heptyl,n-octyl, n-nonyl, n-decyl and the like.

[0369] The term “alkylene” means a divalent saturated hydrocarbon groupwhich may be linear or branched. Unless otherwise defined, such alkylenegroups typically contain from 1 to 10 carbon atoms. Representativealkylene groups include, by way of example, methylene, ethane-1,2-diyl(“ethylene”), propane-1,2-diyl, propane-1,3-diyl, butane-1,4-diyl,pentane-1,5-diyl and the like.

[0370] The term “alkoxy” means a monovalent group of the formula(alkyl)-O—, where alkyl is as defined herein. Representative alkoxygroups include, by way of example, methoxy, ethoxy, n-propoxy,isopropoxy, n-butoxy, sec-butoxy, isobutoxy, tert-butoxy and the like.

[0371] The term “alkenyl” means a monovalent unsaturated hydrocarbongroup which may be linear or branched and which has at least one, andtypically 1, 2 or 3, carbon-carbon double bonds. Unless otherwisedefined, such alkenyl groups typically contain from 2 to 10 carbonatoms. Representative alkenyl groups include, by way of example,ethenyl, n-propenyl, isopropenyl, n-but-2-enyl, n-hex-3-enyl and thelike. The term “alkenylene” means a divalent alkenyl group.

[0372] The term “alkynyl” means a monovalent unsaturated hydrocarbongroup which may be linear or branched and which has at least one, andtypically 1, 2 or 3, carbon-carbon triple bonds. Unless otherwisedefined, such alkynyl groups typically contain from 2 to 10 carbonatoms. Representative alkynyl groups include, by way of example,ethynyl, n-propynyl, n-but-2-ynyl, n-hex-3-ynyl and the like. The term“alkynylene” means a divalent alkynyl group.

[0373] The term “aryl” means a monovalent aromatic hydrocarbon having asingle ring (i.e., phenyl) or fused rings (i.e., naphthalene). Unlessotherwise defined, such aryl groups typically contain from 6 to 10carbon ring atoms. Representative aryl groups include, by way ofexample, phenyl and naphthalene-1-yl, naphthalene-2-yl, and the like.The term “arylene” means a divalent aryl group.

[0374] The term “azacycloalkyl” means a monovalent heterocyclic ringcontaining one nitrogen atom, i.e., a cycloalkyl group in which onecarbon atom has been replaced with a nitrogen atom. Unless otherwisedefined, such azacycloalkyl groups typically contain from 2 to 9 carbonatoms. Representative examples of an azacycloalkyl group arepyrrolidinyl and piperidinyl groups. The term “azacycloalkylene” means adivalent azacycloakyl group. Representative examples of anazacycloalkylene group are pyrrolidinylene and piperidinylene groups.

[0375] The term “cycloalkyl” means a monovalent saturated carbocyclichydrocarbon group. Unless otherwise defined, such cycloalkyl groupstypically contain from 3 to 10 carbon atoms. Representative cycloalkylgroups include, by way of example, cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl and the like. The term “cycloalkylene” means a divalentcycloalkyl group.

[0376] The term “halo” means fluoro, chloro, bromo and iodo.

[0377] The term “heteroaryl” means a monovalent aromatic group having asingle ring or two fused rings and containing in the ring at least oneheteroatom (typically 1 to 3 heteroatoms) selected from nitrogen, oxygenor sulfur. Unless otherwise defined, such heteroaryl groups typicallycontain from 5 to 10 total ring atoms. Representative heteroaryl groupsinclude, by way of example, monovalent species of pyrrole, imidazole,thiazole, oxazole, furan, thiophene, triazole, pyrazole, isoxazole,isothiazole, pyridine, pyrazine, pyridazine, pyrimidine, triazine,indole, benzofuran, benzothiophene, benzimidazole, benzthiazole,quinoline, isoquinoline, quinazoline, quinoxaline and the like, wherethe point of attachment is at any available carbon or nitrogen ringatom. The term “heteroarylene” means a divalent heteroaryl group.

[0378] The term “heterocyclyl” or “heterocyclic” means a monovalentsaturated or unsaturated (non-aromatic) group having a single ring ormultiple condensed rings and containing in the ring at least oneheteroatom (typically 1 to 3 heteroatoms) selected from nitrogen, oxygenor sulfur. Unless otherwise defined, such heterocyclic groups typicallycontain from 2 to 9 total ring carbon atoms. Representative heterocyclicgroups include, by way of example, monovalent species of pyrrolidine,imidazolidine, pyrazolidine, piperidine, 1,4-dioxane, morpholine,thiomorpholine, piperazine, 3-pyrroline and the like, where the point ofattachment is at any available carbon or nitrogen ring atom. The term“heterocyclene” means a divalent heterocyclyl or heterocyclic group.

[0379] The term “divalent hydrocarbon group” means a divalenthydrocarbon group which is composed primarily of carbon and hydrogenatoms and which optionally contains one or more heteroatoms. Suchdivalent hydrocarbon groups may be branched or unbranched, saturated orunsaturated, acyclic or cyclic, aliphatic or aromatic, or combinationsthereof. The divalent hydrocarbon group can optionally containheteroatoms incorporated into the hydrocarbon chain or as substituentsattached to the hydrocarbon chain.

[0380] When a specific number of carbon atoms is intended for aparticular term used herein, the number of carbon atoms is shown inparentheses preceding the term. For example, the term “(1-4C)alkyl”means an alkyl group having from 1 to 4 carbon atoms.

[0381] The term “library” means a collection of more than one compound,i.e., at least two compounds.

[0382] The term “pharmaceutically acceptable salt” means a salt which isacceptable for administration to a patient, such as a mammal (e.g.,salts having acceptable mammalian safety for a given dosage regime).Such salts can be derived from pharmaceutically acceptable inorganic ororganic bases and from pharmaceutically acceptable inorganic or organicacids. Salts derived from pharmaceutically acceptable inorganic basesinclude ammonium, calcium, copper, ferric, ferrous, lithium, magnesium,manganic, manganous, potassium, sodium, zinc and the like. Particularlypreferred are ammonium, calcium, magnesium, potassium and sodium salts.Salts derived from pharmaceutically acceptable organic bases includesalts of primary, secondary and tertiary amines, including substitutedamines, cyclic amines, naturally-occurring amines and the like, such asarginine, betaine, caffeine, choline, N,N′-dibenzylethylenediamine,diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol,ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine,glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine,methylglucamine, morpholine, piperazine, piperadine, polyamine resins,procaine, purines, theobromine, triethylamine, trimethylamine,tripropylamine, tromethamine and the like. Salts derived frompharmaceutically acceptable acids include acetic, ascorbic,benzenesulfonic, benzoic, camphosulfonic, citric, ethanesulfonic,edisylic, fumaric, gentisic, gluconic, glucoronic, glutamic, hippuric,hydrobromic, hydrochloric, isethionic, lactic, lactobionic, maleic,malic, mandelic, methanesulfonic, mucic, naphthalenesulfonic,naphthalene-1,5-disulfonic, naphthalene-2,6-disulfonic, nicotinic,nitric, orotic, pamoic, pantothenic, phosphoric, succinic, sulfuric,tartaric, p-toluenesulfonic, xinafoic and the like. Particularlypreferred are citric, hydrobromic, hydrochloric, isethionic, maleic,naphthalene-1,5-disulfonic, phosphoric, sulfuric and tartaric acids.

[0383] The term “salt thereof” means a compound formed when the hydrogenof an acid is replaced by a cation, such as a metal cation or an organiccation and the like. Preferably, the salt is a pharmaceuticallyacceptable salt, although this is not required for salts of intermediatecompounds that are not intended for administration to a patient.

[0384] The term “solvate” means a complex or aggregate formed by one ormore molecules of a solute, i.e. a compound of formula I or apharmaceutically acceptable salt thereof, and one or more molecules of asolvent. Such solvates are typically crystalline solids having asubstantially fixed molar ratio of solute and solvent. Representativesolvents include, by way of example, water, methanol, ethanol,isopropanol, acetic acid and the like. When the solvent is water, thesolvate formed is a hydrate.

[0385] It will be appreciated that the term “or salt or stereoisomerthereof” is intended to include all permutations of salts andstereoisomers, such as a salt of a stereoisomer of a compound of formulaI.

[0386] The term “therapeutically effective amount” means an amountsufficient to effect treatment when administered to a patient in need oftreatment.

[0387] The term “treating” or “treatment” as used herein means thetreating or treatment of a disease or medical condition (such as COPD)in a patient, such as a mammal (particularly a human) that includes:

[0388] (a) preventing the disease or medical condition from occurring,i.e., prophylactic treatment of a patient;

[0389] (b) ameliorating the disease or medical condition, i.e.,eliminating or causing regression of the disease or medical condition ina patient;

[0390] (c) suppressing the disease or medical condition, i.e., slowingor arresting the development of the disease or medical condition in apatient; or

[0391] (d) alleviating the symptoms of the disease or medical conditionin a patient.

[0392] The term “leaving group” means a functional group or atom whichcan be displaced by another functional group or atom in a substitutionreaction, such as a nucleophilic substitution reaction. By way ofexample, representative leaving groups include chloro, bromo and iodogroups; sulfonic ester groups, such as mesylate, tosylate, brosylate,nosylate and the like; and acyloxy groups, such as acetoxy,trifluoroacetoxy and the like.

[0393] The term “protected derivatives thereof” means a derivative ofthe specified compound in which one or more functional groups of thecompound are protected from undesired reactions with a protecting orblocking group. Functional groups which may be protected include, by wayof example, carboxylic acid groups, amino groups, hydroxyl groups, thiolgroups, carbonyl groups and the like. Representative protecting groupsfor carboxylic acids include esters (such as a p-methoxybenzyl ester),amides and hydrazides; for amino groups, carbamates (such astert-butoxycarbonyl) and amides; for hydroxyl groups, ethers and esters;for thiol groups, thioethers and thioesters; for carbonyl groups,acetals and ketals; and the like. Such protecting groups are well-knownto those skilled in the art and are described, for example, in T. W.Greene and G. M. Wuts, Protecting Groups in Organic Synthesis, ThirdEdition, Wiley, New York, 1999, and references cited therein.

[0394] The term “amino-protecting group” means a protecting groupsuitable for preventing undesired reactions at an amino group.Representative amino-protecting groups include, but are not limited to,tert-butoxycarbonyl (BOC), trityl (Tr), benzyloxycarbonyl (Cbz),9-fluorenylmethoxycarbonyl (Fmoc), formyl, trimethylsilyl (TMS),tert-butyldimethylsilyl (TBS), and the like.

[0395] The term “carboxy-protecting group” means a protecting groupsuitable for preventing undesired reactions at a carboxy group.Representative carboxy-protecting groups include, but are not limitedto, esters, such as methyl, ethyl, tert-butyl, benzyl (Bn),p-methoxybenzyl (PMB), 9-fluroenylmethyl (Fm), trimethylsilyl (TMS),tert-butyldimethylsilyl (TBS), diphenylmethyl (benzhydryl, DPM) and thelike.

[0396] The term “hydroxyl-protecting group” means a protecting groupsuitable for preventing undesirable reactions at a hydroxyl group.Representative hydroxyl-protecting groups include, but are not limitedto, silyl groups including tri(1-6C)alkylsilyl groups, such astrimethylsilyl (TMS), triethylsilyl (TES), tert-butyldimethylsilyl (TBS)and the like; esters (acyl groups) including (1-6C)alkanoyl groups, suchas formyl, acetyl and the like; arylmethyl groups, such as benzyl (Bn),p-methoxybenzyl (PMB), 9-fluorenylmethyl (Fm), diphenylmethyl(benzhydryl, DPM) and the like. Additionally, two hydroxyl groups canalso be protected as an alkylidene group, such as prop-2-ylidine,formed, for example, by reaction with a ketone, such as acetone.

[0397] General Synthetic Procedures for Preparing Library

[0398] The biphenyl derivatives found in the libraries of this inventioncan be prepared from readily available starting materials using thefollowing general methods and procedures or by using other informationreadily available to those of ordinary skill in the art. Although aparticular embodiment of the present invention may be shown or describedherein, those skilled in the art will recognize that all embodiments oraspects of the present invention can be prepared using the methodsdescribed herein or by using other methods, reagents and startingmaterials known to those skilled in the art. It will also be appreciatedthat where typical or preferred process conditions (i.e., reactiontemperatures, times, mole ratios of reactants, solvents, pressures,etc.) are given, other process conditions can also be used unlessotherwise stated. While the optimum reaction conditions may varydepending on the particular reactants or solvent used, such conditionscan be readily determined by one skilled in the art by routineoptimization procedures.

[0399] Additionally, as will be apparent to those skilled in the art,conventional protecting groups may be necessary or desired to preventcertain functional groups from undergoing undesired reactions. Thechoice of a suitable protecting group for a particular functional groupas well as suitable conditions for protection and deprotection of suchfunctional groups are well-known in the art. Protecting groups otherthan those illustrated in the procedures described herein may be used,if desired. For example, numerous protecting groups, and theirintroduction and removal, are described in T. W. Greene and G. M. Wuts,Protecting Groups in Organic Synthesis, Third Edition, Wiley, New York,1999, and references cited therein.

[0400] By way of illustration, the biphenyl derivatives employed in thisinvention can be prepared by a process comprising:

[0401] (a) reacting a compound of formula 1:

[0402] or a salt thereof; with a compound of formula 2:

[0403] wherein X¹ represents a leaving group, and P¹ and P² eachindependently represent a hydrogen atom or a hydroxyl-protecting group;

[0404] (b) reacting a compound of formula 3:

[0405] or salt thereof; with a compound of formula 4:

[0406] wherein X² represents a leaving group, and P³ and P⁴ eachindependently represent a hydrogen atom or a hydroxyl-protecting group;

[0407] (c) coupling a compound of formula 5:

[0408] with a compound of formula 6:

[0409] wherein X^(Qa) and X^(Qb) each independently represent functionalgroups that couple to form a group Q, P^(5a) represents a hydrogen atomor an amino-protecting group; and P^(5b) and P⁶ each independentlyrepresent a hydrogen atom or a hydroxyl-protecting group;

[0410] (d) for a compound of formula I wherein R⁵ represents a hydrogenatom, reacting a compound of formula 3 with a compound of formula 7:

[0411] or a hydrate thereof (e.g., a glyoxal), in the presence of areducing agent, wherein P⁷ represents a hydrogen atom or ahydroxyl-protecting group;

[0412] (e) reacting a compound of formula 1 with a compound of formula8:

[0413] or a hydrate thereof, in the presence of a reducing agent,wherein P⁸ and P⁹ each independently represent a hydrogen atom or ahydroxyl-protecting group, P¹⁰ represents a hydrogen atom or anamino-protecting group, and R^(4′) represents a residue that, togetherwith the carbon to which it is attached, affords a group R⁴ uponcompletion of the reaction;

[0414] (f) reacting a compound of formula 9:

[0415] wherein X³ represents a leaving group, with a compound of formula10:

[0416] wherein P¹¹ and P¹² each independently represent a hydrogen atomor a hydroxyl-protecting group, and P¹³ represents a hydrogen atom or anamino-protecting group; or

[0417] (g) reacting a compound of formula 11:

[0418] or a hydrate thereof; wherein R^(4′) represents a residue that,together with the carbon to which it is attached, affords a group R⁴upon completion of the reaction; with a compound of formula 10 in thepresence of a reducing agent; and then

[0419] (h) removing any protecting group P¹, P², P³, P⁴, P^(5a), P^(5b),P⁶, P⁷, P⁸, P⁹, P¹⁰, P¹¹, P¹² or P¹³ to provide a compound of formula I.A library of compounds of formula I is prepared by repeating steps (a),(b), (c), (d), (e), (f) or (g) and step (h) to prepare each member ofthe library.

[0420] Generally, if a salt of one of the starting materials is used inthe processes described above, such as an acid addition salt, the saltis typically neutralized before or during the reaction process. Thisneutralization reaction is typically accomplished by contacting the saltwith one molar equivalent of a base for each molar equivalent of acidaddition salt.

[0421] In process (a), i.e., the reaction between the compounds offormula 1 and 2, the leaving group represented by X¹ can be, forexample, halo, such as chloro, bromo or iodo, or a sulfonic ester group,such as mesylate or tosylate. The groups P¹ and P² can be, for example,trimethylsilyl and benzyl, respectively. This reaction is typicallyconducted in an inert diluent, such as acetonitrile, in the presence ofa base. For example, this reaction can be conducted in the presence of atertiary amine, such as diisopropylethylamine. Generally, this reactionis conducted at a temperature in the range of from 0° C. to 100° C.until the reaction is substantially complete. The reaction product isthen isolated using conventional procedures, such as extraction,recrystallization, chromatography and the like.

[0422] Compounds of formula 1 are generally known in the art or can beprepared from commercially available starting materials and reagentsusing well-known procedures. For example, compounds of formula 1 can beprepared by deprotecting a compound of formula 12:

[0423] wherein P¹⁴ represents an amino-protecting group, such as abenzyl group. By way of illustration, a benzyl group can be readilyremoved by reduction using, for example, hydrogen or ammonium formateand a group VIII metal catalyst, such as palladium on carbon. When Wrepresents NW^(a), the hydrogenation reaction is conveniently performedusing Pearlman's catalyst (i.e., Pd(OH)₂).

[0424] Compounds of formula 12 can be prepared by reacting an isocyanatecompound of formula 13:

[0425] with a compound of formula 14:

[0426] Compounds of formula 2 can be prepared by various proceduresdescribed herein or by procedures that are well-known to those skilledin the art. For example, the hydroxyl group of a compound of formula 23below, can be readily converted into a leaving group using well-knownreagents and procedures. By way of illustration, a hydroxyl group can beconverted into a halo group using an inorganic acid halide, such asthionyl chloride, phosphorous trichloride, phosphorous tribromide,phosphorous oxychloride and the like, or a halogen acid, such a hydrogenbromide.

[0427] In process (b), i.e., the reaction of a compound of formula 3with a compound of formula 4, the leaving represented by X² can be, forexample, halo, such as chloro, bromo or iodo, or a sulfonic ester group,such as mesylate or tosylate. The groups P³ and P⁴ can be, for example,tert-butyldimethylsilyl and benzyl, respectively. This reaction istypically conducted in the presence of a base, such as sodiumbicarbonate, and an alkali metal iodide, such as sodium iodide.Generally, this reaction is conducted in an inert diluent, such astetrahydrofuran, at a temperature ranging from 25° C. to 100° C. untilthe reaction is substantially complete. The reaction product is thenisolated using conventional procedures, such as extraction,recrystallization, chromatography and the like.

[0428] Compounds of formula 3 can be prepared by deprotecting a compoundof formula 15:

[0429] wherein one or both of P¹⁵ and P¹⁶ independently represents aprotecting group, such as tert-butoxycarbonyl, and any remainderrepresents a hydrogen atom. For example, a tert-butoxycarbonyl group canbe removed by treating the protected compound with trifluoroacetic acid.

[0430] Compounds of formula 15 can be prepared by reacting a compound offormula 1 with a compound of formula 16:

X³—R⁴—NP¹⁵P¹⁶  16

[0431] wherein X³ represents a leaving group such as halo, such aschloro, bromo or iodo, or sulfonic ester group, such as mesylate ortosylate. This reaction is typically conducted by contacting a compoundof formula 1 with a compound of formula 16 in an inert diluent, such asacetonitrile, DMF or mixtures thereof, at a temperature ranging fromabout 0° C. to about 100° C. until the reaction is substantiallycomplete.

[0432] Alternatively, compounds of formula 3 can be obtained byreductive amination of a compound of formula 11. The reductive aminationcan be performed by reacting the compound of formula 11 with, forexample, benzylamine and hydrogen in the presence of palladium oncarbon.

[0433] Compounds of formula 11 may be prepared by oxidizing thecorresponding alcohol of formula 17:

[0434] using a suitable oxidizing agent, such as sulfur trioxidepyridine complex and dimethyl sulfoxide. This oxidation reaction istypically conducted in an inert diluent, such as dichloromethane, thepresence of a tertiary amine, such as diisopropylethylamine, at atemperature ranging from about −20° C. to about 25° C.

[0435] Compounds of formula 17 can be prepared by reacting a compound offormula 1 with a compound of formula 18:

X⁴—R⁴—OH  18

[0436] wherein X⁴ represents a leaving group such as halo, such aschloro, bromo or iodo, or a sulfonic ester group, such as mesylate ortosylate.

[0437] Compounds of formula 4 can be prepared by reacting a compound offormula 19:

[0438] with a reducing agent, such as borane. If desired, such areduction can be performed in the presence of a chiral catalyst toprovide compounds of formula 4 in chiral form. For example, compounds offormula 19 can be reduced in the presence of a chiral catalyst formedfrom (R)-(+)-α,α-diphenyl-2-pyrrolidinemethanol and trimethylboroxine;or alternatively, from (S)-(−)-α,α-diphenyl-2-pyrrolidinemethanol andtrimethylboroxine. The resulting hydroxyl group can then be protectedwith a hydroxyl-protecting group, P³, by reaction with, for example,tert-butyldimethylsilyl trifluoromethanesulfonate.

[0439] Compounds of formula 19 in which X² represents a bromine atom canbe prepared by reacting a compound of formula 20:

[0440] with bromine in the presence of a Lewis acid, such as borontrifluoride diethyl etherate. Compounds of formula 20 are well-known inthe art or can be prepared by well-known procedures using commerciallyavailable starting materials and reagents.

[0441] Referring to process (c), i.e., the reaction of a compound offormula 5 with a compound of formula 6, it will be appreciated that thegroups X^(Qa) and X^(Qb) should be selected so as to afford the desiredgroup Q upon completion of the reaction. For example, when the desiredgroup Q is an amide group, i.e., —N(Q^(a))C(O)— or —C(O)N(Q^(b)), one ofX^(Qa) and X^(Qb) can be an amine group (i.e., —NHQ^(a) or —NHQ^(b)) andthe other can be a carboxyl group (i.e., —COOH) or a reactive derivativethereof (such as acyl halide, such as an acyl chloride or acyl bromide).The groups P^(5a), P^(5b) and P⁶ can be, for example, benzyl,trimethylsilyl and benzyl, respectively. When Q is an amide group, thereaction can be performed under conventional amide coupling conditions.Similarly, when the desired group Q is a sulfonamide, i.e.,—N(Q^(c))S(O)₂— or —S(O)₂N(Q^(d))—, one of X^(Qa) and X^(Qb) can be anamine group, —NHQ^(c) or —NHQ^(d) and the other can be a sulfonyl halidegroup (such as sulfonyl chloride or sulfonyl bromide).

[0442] Compounds of formula 5 can be prepared by reacting a compound offormula 1 with a compound of formula 21:

X⁵—(R^(4a))_(d)—(A¹)_(e)—(R_(4b))_(f)—X^(Qa′)  21

[0443] wherein X⁵ represents a leaving group including halo, such aschloro, bromo or iodo, and a sulfonic ester group, such as mesylate ortosylate; and X^(Qa′) represents X^(Qa), such as a carboxyl group or anamino group NHQ^(a), or a protected derivative thereof, such as a(1-6C)alkoxycarbonylamino group or a tert-butoxycarbonylamino group.This reaction is typically conducted by a method analogous to that usedto prepare compounds of formula 3, followed by removing any protectinggroup in X^(Qa′).

[0444] Compounds of formula 6 can be prepared by reacting a compound offormula 4 with a compound of formula 22:

X^(Qb′)—(R^(4c))_(g)—(A²)_(h)—(R^(4d))_(i)—X⁶  22

[0445] wherein X⁶ represents a leaving group including halo, such aschloro, bromo or iodo, and a sulfonic ester group, such as mesylate ortosylate; and X^(Qb′) represents X^(Qb), such as a carboxyl group or anamino group NHQ^(b), or a protected derivative thereof, such as a(1-6C)alkoxycarbonyl group or a tert-butoxycarbonylamino group. Thisreaction is typically conducted by a method analogous to that used toprepare compounds of formula 3, followed by removing any protectinggroup in X^(Qb′).

[0446] Referring to process (d), i.e., the reaction of a compound offormula 3 with a compound of formula 7, any suitable reducing agent maybe used in this reaction. For example, the reducing agent can behydrogen in the presence of a Group VIII metal catalyst, such aspalladium on carbon; or a metal hydride reagent, such as sodiumtriacetoxyborohydride. The group P⁷ can be, for example, benzyl. Thisreaction is typically conducted in an inert diluent and a proticsolvent, such as a mixture of dichloroethane and methanol, at atemperature in the range of from 0° C. to 100° C. until the reaction issubstantially complete.

[0447] Compounds of formula 7 in the form of a hydrate can be preparedby conventional procedures, for example, by dibrominating a compound offormula 19 (where X² in this case can also be hydrogen), and thenhydrolyzing the resulting dibromide to form a glyoxal or a hydratethereof. For example, a compound of formula 19 can be reacted withhydrogen bromide and then hydrolyzed with water to form thecorresponding glyoxal hydrate.

[0448] Referring to process (e), i.e., the reaction of a compound offormula 1 with a compound of formula 8, any suitable reducing agent maybe used in this reaction. For example, the reducing agent may behydrogen in the presence of a Group VIII metal catalyst, such aspalladium on carbon; or a metal hydride reagent, such as sodiumtriacetoxyborohydride. The groups P⁸, P⁹ and P¹⁰ can be, for example,trimethylsilyl, benzyl and benzyl, respectively. Typically, thisreduction reaction is conducted in an inert diluent and a proticsolvent, such as dichloroethane and methanol, at a temperature in therange of from 0° C. to 100° C. until the reaction is substantiallycomplete.

[0449] Compounds of formula 8 may be prepared by oxidizing a compound offormula 23:

[0450] using any suitable oxidizing agent, such as sulfur trioxidepyridine complex and dimethyl sulfoxide. This reaction is typicallyconducted in the presence of a tertiary amine, such asdiisopropylethylamine, at a temperature in the range of from about −20°C. to about 25° C. until the oxidation is substantially complete.

[0451] Compounds of formula 23 can be prepared by reacting a compound offormula 10 with a compound of formula 24:

HO—R⁴—X⁷  24

[0452] wherein X⁷ represents a leaving group including halo, such aschloro, bromo or iodo, and a sulfonic ester group, such as mesylate ortosylate.

[0453] Referring to process (f), i.e., the reaction of a compound offormula 9 with a compound of formula 10, the leaving group representedby X³ can be, for example, halo, such as chloro, bromo or iodo, or asulfonic ester group, such as mesylate or tosylate. The groups P¹¹, P¹²and P¹³ can be, for example, trimethylsilyl, benzyl and benzyl,respectively. This reaction is typically conducted an inert diluent,such as acetonitrile, in the presence of a suitable base. For example,this reaction can be conducted in the presence of a tertiary amine, suchas diisopropylethylamine. Generally, this reaction is conducted at atemperature in the range of from 0° C. to 100° C. until the reaction issubstantially complete.

[0454] Compounds of formula 9 can be prepared by steps analogous tothose of methods (a) to (e) herein, starting from a compound offormula 1. Additionally, compounds of formula 10 can be prepared fromcompounds of formula 4 by reaction with an amine of formula P¹³NH₂.

[0455] Referring to process (g), i.e., the reaction of a compound offormula 11 with a compound of formula 10, any suitable reducing agentmay be used in this reaction. For example, the reducing agent may behydrogen in the presence of a Group VIII metal catalyst, such aspalladium on carbon; or a metal hydride reagent, such as sodiumtriacetoxyborohydride. The groups P¹¹, P¹² and P¹³ can be, for example,tert-butyldimethylsilyl, benzyl and benzyl, respectively. Typically,this reduction reaction is conducted in an inert diluent and a proticsolvent, such as dichloroethane and methanol, at a temperature in therange of from 0° C. to 100° C. until the reaction is substantiallycomplete.

[0456] Compounds of formula 11 are readily prepared by oxidation of thecorresponding alcohol or by hydrolysis of the corresponding acetal. Anysuitable oxidizing agent may be employed in this reaction to provide thealdehyde, such as sulfur trioxide pyridine complex and dimethylsulfoxide. The acetal may be hydrolyzed under conventional conditionsusing aqueous acid to provide the aldehyde.

[0457] In a particular embodiment, certain compounds of formula I areprepared by a process comprising:

[0458] (h) deprotecting a compound of formula 25:

[0459] wherein P¹⁷ represents a hydrogen atom or an amino-protectinggroup; and each of P¹⁸, P¹⁹ and P²⁰ independently represent a hydrogenatom or a hydroxyl-protecting group; provided that at least one of P¹⁷,P¹⁸, P¹⁹ or P²⁰ is a protecting group;

[0460] (i) deprotecting a compound of formula 26:

[0461] wherein P²¹ represents a hydrogen atom or an amino-protectinggroup; and each of P²² and P²³ independently represent a hydrogen atomor a hydroxyl-protecting group; provided that at least one of P²¹, P²²or P²³ is a protecting group; or

[0462] (j) deprotecting a compound of formula 27:

[0463] wherein P²⁴ represents a hydrogen atom or an amino-protectinggroup; and each of P²⁵ and P²⁶ independently represent a hydrogen atomor a hydroxyl-protecting group; provided that at least one of P²⁴, P²⁵or P²⁶ is a protecting group.

[0464] to provide a compound of formula I.

[0465] Referring to process (h), examples of particular values for P¹⁷,P¹⁸, P¹⁹ and P²⁰ are: for P¹⁷, hydrogen or benzyl; for P¹⁸ hydrogen ortert-butyldimethylsilyl; and for P¹⁹ and P²⁰ hydrogen or benzyl, ortogether propylidine. In this process, benzyl protecting groups areconveniently removed by catalytic hydrogenation in the presence of aGroup VIII metal catalyst, such as palladium on carbon; atert-butyldimethylsilyl group is conveniently removed by treatment withhydrogen fluoride, such as triethylamine trihydrofluoride; and apropylidine group is conveniently removed by treatment with an acid,such as trifluoroacetic acid.

[0466] Compounds of formula 25 can be prepared by the methods describedherein, such as by processes (a) to (g). Alternatively, compounds offormula 25 can be prepared by reacting a compound of formula 28:

[0467] wherein R⁸ represents —CH₂OP¹⁹, —CHO, —COOH or—C(O)O(1-6C)alkoxy, such as carbomethoxy, R⁹ represents —OP¹⁸ and R¹⁰represents a hydrogen atom, or R⁹ and R¹⁰ together represent ═O, with areducing agent. Any suitable reducing agent may be used in this reactionincluding, by way of example, metal hydride reducing agents, such assodium borohydride, lithium aluminum hydride and the like.

[0468] Compounds of formula 28 in which R⁹ and R¹⁰ together represent a═O group can be readily prepared by reacting a compound of formula 29:

[0469] or a salt thereof, with a compound of formula 30:

[0470] wherein X⁸ represents a leaving group, such as a bromo.

[0471] Referring to process (i), examples of particular values for P²¹,P²² and P²³ are: for P²¹, hydrogen or benzyl; for P²² hydrogen ortert-butyldimethylsilyl; and for P²³ hydrogen or benzyl. In thisprocess, benzyl protecting groups are conveniently removed by catalytichydrogenation in the presence of a Group VIII metal catalyst, such aspalladium on carbon; and a tert-butyldimethylsilyl group is convenientlyremoved by treatment with hydrogen fluoride, such as triethylaminetrihydrofluoride. Compounds of formula 26 can be prepared by the methodsdescribed herein, such as by processes (a) to (g).

[0472] Referring to process (j), examples of particular values for P²⁴,P²⁵ and P²⁶ are: for P²⁴, hydrogen or benzyl; for P²⁵ hydrogen ortert-butyldimethylsilyl; and for P²⁶ hydrogen or benzyl. In thisprocess, benzyl protecting groups are conveniently removed by catalytichydrogenation in the presence of a Group VIII metal catalyst, such aspalladium on carbon; and a tert-butyldimethylsilyl group is convenientlyremoved by treatment with hydrogen fluoride, such as triethylaminetrihydrofluoride. Compounds of formula 27 can be prepared by the methodsdescribed herein, such as by processes (a) to (g).

[0473] Additionally, compounds of formula I in which R⁶ and R⁷ togetherform —NR^(7g)C(O)—CR^(7h)R^(7i)—CR^(7j)R^(7k)— or—CR^(7l)R^(7m)—CR^(7n)R^(7o)—C(O)—NR^(7p)— may be prepared by reducing acorresponding compound of formula I in which R⁶ and R⁷ together form—NR^(7a)C(O)—CR^(7b)═CR^(7c)— or —CR^(7d)═CR^(7e)—C(O)—NR^(7f)—, forexample by catalytic hydrogenation as described in Example 6hereinafter.

[0474] Further details regarding specific reaction conditions and otherprocedures for preparing members of the libraries of this invention orintermediates thereof are described in the Examples set forth below.

[0475] Libraries of Biphenyl Derivatives

[0476] The libraries of this invention typically contain at least twocompounds of formula I. For example, in one embodiment, the librarycontains from 2 to 1,000 compounds; including, by way of illustrationfrom 2 to 500 compounds; or from 2 to 200 compounds; or from 2 to 100compounds; or from 2 to 50 compounds. If desired, the library cancontain more than 1000 compounds, such as up to 5,000 or 10,000compounds or more.

[0477] The members of the library can be synthesized individually, or inarrays or as mixtures. Typically, each compound of the library will beprepared individually in a separate reaction vessel or as a member of anarray, i.e. in a separate well of a multi-well plate. Methods forpreparing compounds of formula I are described in further detail in theExamples hereinbelow.

[0478] Once a library of compounds of formula I has been prepared, themembers of the library are evaluated or screened to determine whether acompound in the library has both β₂ adrenergic receptor agonist andmuscarinic receptor antagonist activity. Any suitable test, assay orprocedure may be used to evaluate the members of the library, includingnumerous in vitro and in vivo assays which are well-known to thoseskilled in the art. Generally, all members of the library will beevaluated, however, if desired, the evaluation process may be terminatedat any point once a compound having both β₂ adrenergic receptor agonistand muscarinic receptor antagonist activity has been found.

[0479] By way of illustration, suitable in vitro and in vivo assays caninclude radioligand binding assays, functional assays, bronchoprotectionanimal models and the like. For example, suitable functional assaysinclude ligand-mediated changes in intracellular cyclic adenosinemonophosphate (cAMP), ligand-mediated changes in activity of the enzymeadenylyl cyclase (which synthesizes cAMP), ligand-mediated changes inincorporation of guanosine 5′-O-(-thio)triphosphate ([³⁵S]GTP S) intoisolated membranes via receptor catalyzed exchange of [³⁵S]GTP S forGDP, ligand-mediated changes in free intracellular calcium ions(measured, for example, with a fluorescence-linked imaging plate readeror FLIPR® from Molecular Devices, Inc.). By way of further example,assays suitable for evaluating compounds of formula I are described indetail in the Examples hereinbelow.

[0480] Utility

[0481] The libraries and methods of this invention are useful foridentifying compounds having both β₂ adrenergic receptor agonist andmuscarinic receptor antagonist activity. Compounds having both β₂adrenergic receptor agonist and muscarinic receptor antagonist activityare useful for treating medical conditions mediated by β₂ adrenergicreceptors or muscarinic receptors, i.e., medical conditions that areameliorated by treatment with a β₂ adrenergic receptor agonist or amuscarinic receptor antagonist. Such medical conditions include, by wayof example, pulmonary disorders or diseases associated with reversibleairway obstruction, such as chronic obstructive pulmonary disease (e.g.,chronic and wheezy bronchitis and emphysema), asthma, pulmonary fibrosisand the like. Other conditions which may be treated include prematurelabor, depression, congestive heart failure, skin diseases (e.g.,inflammatory, allergic, psoriatic and proliferative skin diseases,conditions where lowering peptic acidity is desirable (e.g., peptic andgastric ulceration) and muscle wasting disease.

[0482] Among other properties, compounds of particular interest that canbe identified using the libraries and methods of this invention arethose that demonstrate a K_(i) value of less than about 300 nM for a β₂adrenergic receptor and a K_(i) value less than 300 nM for a muscarinicreceptor.

[0483] Another group of compounds of particular interest that can beidentified using the libraries and methods of this invention are thosethat demonstrate an inhibitory constant K_(i) value for binding at amuscarinic receptor, such as a M₃ muscarinic receptor, of less thanabout 100 nM; particularly less than 10 nM; and an EC₅₀ value for β₂adrenergic receptor agonist activity of less than about 100 nM;particularly less than 10 nM. Among these compounds, compounds ofspecial interest include those having muscarinic activity, expressed interms of the inhibitory constant K_(i) for binding at the M₃ muscarinicreceptor, that is about equal to the compound's β₂ adrenergic agonistactivity, expressed in terms of the half maximal effective concentrationEC₅₀, as determined in the in vitro assays described herein, or insimilar assays. For example, compounds of particular interest are thosehaving a ratio of the inhibitory constant K_(i) for the M₃ muscarinicreceptor to the EC₅₀ for the β₂ adrenergic receptor ranging from about30:1 to about 1:30; including about 20:1 to about 1:20; such as about10:1 to about 1:10.

[0484] Additionally, those skilled in the art will recognize that, evenif a particular library does not contain any compound having both β₂adrenergic receptor agonist and muscarinic receptor antagonist activity,such information is useful for designing new libraries to be screenedfor bifunctional activity.

[0485] Representative examples of the libraries and methods of thisinvention are demonstrated by the following synthetic procedures and invitro and in vivo assays.

Examples

[0486] The following Preparations and Examples are provided toillustrate specific embodiments of this invention. These specificembodiments, however, are not intended to limit the scope of thisinvention in any way unless specifically indicated.

[0487] The following abbreviations have the following meanings unlessotherwise indicated and any other abbreviations used herein and notdefined have their standard meaning:

[0488] AC adenylyl cyclase

[0489] Ach acetylcholine

[0490] ATCC American Type Culture Collection

[0491] BSA bovine serum albumin

[0492] cAMP 3′-5′ cyclic adenosine monophosphate

[0493] CHO Chinese hamster ovary

[0494] cM₅ cloned chimpanzee M₅ receptor

[0495] DCM dichloromethane (i.e., methylene chloride)

[0496] DIPEA N,N-diisopropylethylamine

[0497] dPBS Dulbecco's phosphate buffered saline

[0498] DMEM Dulbecco's Modified Eagle's Medium

[0499] DMSO dimethyl sulfoxide

[0500] EDTA ethylenediaminetetraacetic acid

[0501] Emax maximal efficacy

[0502] EtOAc ethyl acetate

[0503] EtOH ethanol

[0504] FBS fetal bovine serum

[0505] FLIPR fluorometric imaging plate reader

[0506] Gly glycine

[0507] HATU O-(7-azabenzotriazol-1-yl-N,N′,N′-tetramethyluroniumhexafluorophosphate

[0508] HBSS Hank's buffered salt solution

[0509] HEK human embryonic kidney cells

[0510] HEPES 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid

[0511] hM₁ cloned human M₁ receptor

[0512] hM₂ cloned human M₂ receptor

[0513] hM₃ cloned human M₃ receptor

[0514] hM₄ cloned human M₄ receptor

[0515] hM₅ cloned human M₅ receptor

[0516] HPLC high-performance liquid chromatography

[0517] IBMX 3-isobutyl-1-methylxanthine

[0518] % Eff % efficacy

[0519] PBS phosphate buffered saline

[0520] PyBOP benzotriazol-1-yloxytripyrrolidinophosphoniumhexafluorophosphate

[0521] rpm rotations per minute

[0522] TFA trifluoroacetic acid

[0523] THF tetrahydrofuran

[0524] Tris tris(hydroxymethyl)aminomethane

[0525] Unless noted otherwise, reagents, starting materials and solventswere purchased from commercial suppliers (such as Aldrich, Fluka, Sigmaand the like) and were used without further purification.

[0526] In the examples described below, HPLC analysis was conductedusing an Agilent (Palo Alto, Calif.) Series 1100 instrument with ZorbaxBonus RP 2.1×50 mm columns, supplied by Agilent, (a C14 column), havinga 3.5 micron particle size. Detection was by UV absorbance at 214 nm.HPLC 10-70 data was obtained with a flow rate of 0.5 mL/minute of10%-70% B over 6 minutes. Mobile phase A was 2%-98%-0.1% ACN-H₂O-TFA;and mobile phase B was 90%-10%-0.1% ACN-H₂O-TFA. Using the mobile phasesA and B described above, HPLC 5-35 data and HPLC 10-90 data wereobtained with a 5 minute gradient.

[0527] Liquid chromatography mass spectrometry (LCMS) data were obtainedwith an Applied Biosystems (Foster City, Calif.) model API-150EXinstrument. LCMS 10-90 data was obtained with a 10%-90% mobile phase Bover a 5 minute gradient.

[0528] Small scale purification was conducted using an API 150EX PrepWorkstation system from Applied Biosystems. The mobile phase was A:water+0.05% v/v TFA; and B: acetonitrile+0.05% v/v TFA. For arrays(typically about 3 to 50 mg recovered sample size) the followingconditions were used: 20 mL/min flow rate; 15 min gradients and a 20mm×50 mm Prism RP column with 5 micron particles (ThermoHypersil-Keystone, Bellefonte, Pa.). For larger scale purifications(typically greater than 100 mg crude sample), the following conditionswere used: 60 mL/min flow rate; 30 min gradients and a 41.4 mm×250 mmMicrosorb BDS column with 10 micron particles (Varian, Palo Alto,Calif.).

[0529] The specific rotation for chiral compounds (indicated as [α]²⁰_(D)) was measured using a Jasco Polarimeter (Model P-1010) with atungsten halogen light source and a 589 nm filter at 20° C. Samples oftest compounds were typically measured at 1 mg/mL water.

[0530] A library of compounds of formula I (i.e., Examples 1-170 and180-186) was prepared as follows:

Preparation 1 N-1,1′-Biphenyl-2-yl-N′-4-(1-benzyl)piperidinylurea

[0531] Biphenyl-2-isocyanate (50 g, 256 mmol) was dissolved inacetonitrile (400 mL) at ambient temperature. After cooling to 0° C., asolution of 4-amino-N-benzylpiperidine (48.8 g, 256 mmol) inacetonitrile (400 mL) was added over 5 min. A precipitate was observedimmediately. After 15 min, acetonitrile (600 mL) was added, and theresultant viscous mixture was stirred for 12 h at 35° C. The solids werethen filtered off and washed with cold acetonitrile, then dried undervacuum, yielding the title compound (100 g, 98% yield). MS m/z: [M+H⁺]calcd for C₂₅H₂₇N₃O 386.22; found 386.3.

Preparation 2 N-1,1′-Biphenyl-2-yl-N′-4-piperidinylurea

[0532] The product of Preparation 1 (20 g, 52 mmol) was dissolved in amixture of anhydrous methanol and anhydrous DMF (3:1, 800 mL). Aqueoushydrochloric acid (0.75 mL of 37% conc. solution, 7.6 mmol) was addedand nitrogen gas was bubbled through the solution vigorously for 20 min.Pearlman's catalyst (Pd(OH)₂, 5 g) was added under a stream of nitrogen,before placing the reaction mixture under a hydrogen atmosphere(balloon). The reaction mixture was allowed to stir for 4 days and wasthen passed twice through pads of Celite to remove the catalyst. Thesolvent was then removed under reduced pressure to yield the titlecompound (13 g, 85% yield). MS m/z: [M+H⁺] calcd for C₁₈H₂₁N₃O 296.17;found 296.0.

[0533] Alternatively, N-1,1′-biphenyl-2-yl-N′-4-piperidinylurea wassynthesized by heating together biphenyl-2-isocyanate (50 g, 256 mmol)and 4-amino-N-benzylpiperidine (51.1 g, 269 mmol) at 70° C. for 12 h(the reaction was monitored by LCMS). The reaction mixture was cooled to50° C. and ethanol (500 mL) added, followed by slow addition of 6Mhydrochloric acid (95 mL). The reaction mixture was cooled to roomtemperature. Ammonium formate (48.4 g, 768 mmol) was added to thereaction mixture and nitrogen gas bubbled through the solutionvigorously for 20 min, before adding palladium (10 wt. % (dry basis) onactivated carbon) (10 g). The reaction mixture was heated at 40° C. for12 h, and then filtered through a pad of Celite and the solvent wasremoved under reduced pressure. To the crude residue was added 1Mhydrochloric acid (20 mL) and 10N sodium hydroxide was added to adjustthe pH to 12. The aqueous layer was extracted with ethyl acetate (2×80mL), dried (magnesium sulfate) and solvent removed under reducedpressure to give the title compound as a solid (71.7 g, 95% yield). MSm/z: [M+H⁺] calcd for C₁₈H₂₁N₃O 296.17; found 296.0.

Preparation 3N-1,1′-Biphenyl-2-yl-N′-4-[1-(9-hydroxynonyl)]piperidinylurea

[0534] 9-Bromo-1-nonanol (4.84 g, 21.7 mmol) was added to a stirredsolution of the product of Preparation 2 (5.8 g, 19.7 mmol) anddiisopropylethylamine (10.29 mL, 59.1 mmol) in acetonitrile (99 mL) at50° C. The reaction mixture was heated at 50° C. for 8 h. The reactionmixture was then allowed to cool and the solvent was removed underreduced pressure. The residue was dissolved in dichloromethane (100 mL),washed with saturated aqueous sodium bicarbonate (2×50 mL) and dried(magnesium sulfate). The solvent was removed under reduced pressure. Thecrude product was purified by flash chromatography(dichloromethane:methanol:ammonia system) to yield the title compound(7.1 g, 16.2 mmol, 82% yield).

Preparation 4 N-1,1′-Biphenyl-2-yl-N′-4-[1-(9-oxononyl)]piperidinylurea

[0535] Dimethyl sulfoxide (490 μL, 6.9 mmol), followed bydiisopropylethylamine (324 μL, 3.45 mmol) was added to a solution of theproduct of Preparation 3 (500 mg, 1.15 mmol) in dichloromethane (11.5mL) at −10° C. under an atmosphere of nitrogen. The reaction mixture wasstirred at −15° C. for 15 min, and then sulfur trioxide pyridine complexwas added portionwise (549 mg, 3.45 mmol). The reaction mixture wasstirred at −15° C. for 1 h, and then water (10 mL) was added. Theorganic phase was then separated, washed with water (10 mL), and dried(sodium sulfate). The solvent was removed under reduced pressure to givethe title compound (475 mg, 1.09 mmol, 95% yield). HPLC (10-70)R_(t)=3.39.

Preparation 5N-1,1′-Biphenyl-2-yl-N′-4-[1-(9-aminononyl)]piperidinylurea

[0536] Palladium (10 wt. % (dry basis) on activated carbon) (1.5 g) wasadded to a stirred solution of the product of Preparation 4 (1.58 g,3.63 mmol) and benzylamine (516 μL, 4.72 mmol) in methanol (36.3 mL).The reaction mixture was placed under an atmosphere of hydrogen. Afterstirring for 12 h, the reaction mixture was filtered through a pad ofCelite and washed with methanol (10 mL). The solvent was removed underreduced pressure to give the title compound (1.50 g, 3.45 mmol, 95%yield). HPLC (10-70) R_(t)=2.35; MS m/z: [M+H⁺] calcd for C₂₇H₄₀N₄O₁437.06; found 437.5.

Preparation 6 8-Benzyloxy-5-(2,2-dihydroxyacetyl)-1H-quinolin-2-one

[0537] (a) 8-Acetoxy-1H-quinolin-2-one

[0538] 8-Hydroxyquinoline-N-oxide (160.0 g, 1.0 mol),commercially-available from Aldrich, Milwaukee, Wis., and aceticanhydride (800 mL, 8.4 mol) were heated at 100° C. for 3 h and thencooled in ice. The product was collected on a Buchner funnel, washedwith acetic anhydride (2×100 mL) and dried under reduced pressure togive 8-acetoxy-1H-quinolin-2-one (144 g) as a solid.

[0539] (b) 5-Acetyl-8-hydroxy-1H-quinolin-2-one

[0540] A slurry of aluminum chloride (85.7 g, 640 mmol) in1,2-dichloroethane (280 mL) was cooled in ice, and the product of step(a) (56.8 g, 280 mmol) was added. The mixture was warmed to roomtemperature and then heated at 85° C. After 30 min, acetyl chloride (1.5mL, 21 mmol) was added and the mixture was heated an additional 60 min.The reaction mixture was then cooled and added to 1N hydrochloric acid(3 L) at 0° C. with good stirring. After stirring for 2 h, the solidswere collected on a Buchner funnel, washed with water (3×250 mL) anddried under reduced pressure. The crude product isolated from severalbatches (135 g) was combined and triturated with dichloromethane (4 L)for 6 h. The product was collected on a Buchner funnel and dried underreduced pressure to give 5-acetyl-8-hydroxy-2(1H)-quinolinone (121 g).

[0541] (c) 5-Acetyl-8-benzyloxy-1H-quinolin-2-one

[0542] To the product of step (b) (37.7 g, 186 mmol) was addedN,N-dimethylformamide (200 mL) and potassium carbonate (34.5 g, 250mmol) followed by benzyl bromide (31.8 g, 186 mmol). The mixture wasstirred at room temperature for 2.25 hour and then poured into saturatedsodium chloride (3.5 L) at 0° C. and stirred for 1 hour. The product wascollected and dried on a Buchner funnel for 1 hour, and the resultingsolids were dissolved in dichloromethane (2 L) and this mixture wasdried over sodium sulfate. The solution was filtered through a pad ofCelite which was then washed with dichloromethane (5×200 mL). Thecombined filtrate was then concentrated to dryness and the resultingsolids were triturated with ether (500 mL) for 2 h. The product wascollected on a Buchner funnel, washed with ether (2×250 mL) and driedunder reduced pressure to give 5-acetyl-8-benzyloxy-1H-quinolin-2-one(44 g) as a powder.

[0543] (d) 8-Benzyloxy-5-(2,2-dihydroxyacetyl)-1H-quinolin-2-one

[0544] To a slurry of the product of step (c) (10.0 g, 34.1 mmol) inDMSO (60 mL) was added a 48% w/w hydrobromic acid solution (11.8 mL,102.3 mmol). The mixture was warmed to 60° C. for 16 h then allowed tocool to room temperature. Water (100 mL) was added and the resultingslurry stirred at room temperature for 0.5 h before being cooled to 0°C. The product was collected on a Buchner funnel then dried underreduced pressure to give8-benzyloxy-5-(2,2-dihydroxyacetyl)-1H-quinolin-2-one (12.2 g) as asolid.

Preparation 71-(1-{9-[2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethylamino]nonyl}piperidin-4-yl)-3-biphenyl-2-ylurea

[0545] The products of Preparation 5 (183 mg, 0.42 mmol) and Preparation6 (149 mg, 0.46 mmol) were stirred in dichloroethane (4.2 mL) at ambienttemperature for 2 h. Sodium triacetoxyborohydride (267 mg, 1.26 mmol)was then added and the reaction mixture was stirred for a further 12 h.The reaction mixture was then diluted with dichloromethane (10 mL),washed with saturated aqueous sodium bicarbonate (10 mL), dried(magnesium sulfate) and the solvent was removed under reduced pressure.The crude reaction mixture was purified by flash chromatography (5-10%methanol in dichloromethane, 0.5% ammonium hydroxide) to give the titlecompound (144 mg, 0.20 mmol, 48% yield). HPLC (10-70) R_(t)=3.48; MSm/z: [M+H⁺] calcd for C₄₅H₅₅N₅O₄ 730.4; found 730.7.

Example 11-Biphenyl-2-yl-3-(1-{9-[2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]nonyl}piperidin-4-yl)urea

[0546] Palladium (10 wt. % (dry basis) on activated carbon) (63 mg) wasadded to a stirred solution of the product of Preparation 7 (144 mg,0.20 mmol) in methanol (2 mL) and the reaction mixture was placed underan atmosphere of hydrogen. After 12 h stirring, the reaction mixture wasfiltered through a pad of Celite, washed with methanol (2 mL) and thenthe solvent was removed under reduced pressure. The resulting residuewas purified by preparative HPLC to give the title compound (10 mg).HPLC (10-70) R_(t)=2.8; MS m/z: [M+H⁺] calcd for C₃₈H₄₉N₅O₄ 640.3; found640.5.

Preparation 8 Biphenyl-2-ylcarbamic Acid Piperidin-4-yl Ester

[0547] Biphenyl-2-isocyanate (97.5 g, 521 mmol) and4-hydroxy-1-benzylpiperidine (105 g, 549 mmol), bothcommercially-available from Aldrich, Milwaukee, Wis., were heatedtogether at 70° C. for 12 h, during which time the formation ofbiphenyl-2-ylcarbamic acid 1-benzylpiperidin-4-yl ester was monitored byLCMS. The reaction mixture was then cooled to 50° C. and ethanol (1 L)was added, and then 6M hydrochloric acid (191 mL) was added slowly. Thereaction mixture was then cooled to ambient temperature and ammoniumformate (98.5 g, 1.56 mol) was added and nitrogen gas was bubbledthrough the solution vigorously for 20 min. Palladium (10 wt. % (drybasis) on activated carbon) (20 g) was then added. The reaction mixturewas heated at 40° C. for 12 h and then filtered through a pad of Celite.The solvent was then removed under reduced pressure and 1M hydrochloricacid (40 mL) was added to the crude residue. Sodium hydroxide (10N) wasthen added to adjust the pH to 12. The aqueous layer was extracted withethyl acetate (2×150 mL) and dried (magnesium sulfate), and then thesolvent was removed under reduced pressure to give the title compound(155 g, 100%). HPLC (10-70) R_(t)=2.52; MS m/z: [M+H⁺] calc'd forC₁₈H₂₀N₂O₂ 297.15; found 297.3.

Preparation 9 N,N-(Di-tert-butoxycarbonyl)-9-bromononylamine

[0548] A solution of di-tert-butoxycarbonylamine (3.15 g, 14.5 mmol) inN,N-dimethylformamide (0.28 mL) was cooled to 0° C. for about 10 min.Sodium hydride, 60% in mineral oil (0.58 g, 14.5 mmol) was added and thereaction mixture was stirred at 0° C. for 10 min. The reaction mixturewas removed from the ice bath and allowed to warm to room temperaturefor about 30 min. The reaction mixture was then cooled back down to 0°C. and a solution of 1,9-dibromononane (2.46 mL, 12.1 mmol) indimethylformamide (100 mL) was added. The reaction mixture was stirredovernight at room temperature. After 24 h, MS analysis showed that thereaction was completed. The reaction mixture was concentrated to drynessand diluted with ethyl acetate (100 mL). The organic layer was washedwith saturated sodium bicarbonate (2×100 mL), brine (100 mL), dried(magnesium sulfate) and concentrated under reduced pressure to yield thecrude product, which was purified by chromatography on silica gel using5% ethyl acetate in hexanes to afford the title compound. MS m/z: [M+H⁺]calcd for C₁₉H₃₆N₁O₄Br 423.18; found 423.

Preparation 10 Biphenyl-2-ylcarbamic Acid1-(9-Di-tert-butoxycarbonylamino)nonyl]piperidin-4-yl Ester

[0549] A mixture of 1:1 acetonitrile and N,N-dimethylformamide (50 mL)was added to the products of Preparation 8 (3.0 g, 10.1 mmol) andPreparation 9 (5.1 g, 12.2 mmol) and triethylamine (1.42 mL, 10.1 mmol).The reaction mixture was stirred at ambient temperature for 24 h and wasmonitored by LCMS analysis. The reaction mixture was then concentratedand diluted with ethyl acetate (50 mL). The organic layer was washedwith saturated sodium bicarbonate (2×50 mL) and brine (50 mL). Theorganic phase was then dried over magnesium sulfate and concentrated toyield 6.5 g of crude oil. The oil was purified by chromatography onsilica gel using 1:1 hexanes/ethyl acetate to provide the title compound(3 g). MS m/z: [M+H⁺] calcd for C₃₇H₅₅N₃O₆ 638.41; found 639.

Preparation 11 Biphenyl-2-ylcarbamic Acid 1-(9-Aminononyl)piperidin-4-ylEster

[0550] Trifluoroacetic acid (11 mL) was added to a solution of theproduct of Preparation 10 (7.2 g, 11.3 mmol) in dichloromethane (56 mL).After 2 h, LCMS analysis showed that the reaction was completed. Thereaction mixture was then concentrated to dryness and diluted with ethylacetate (75 mL). Sodium hydroxide (IN) was then added until the pH ofthe mixture reached 14. The organic phase was then collected and washedwith saturated sodium bicarbonate (2×50 mL) and brine (50 mL). Theorganic phase was then dried over magnesium sulfate and concentrated toprovide the title compound (5.5 g). MS m/z: [M+H⁺] calcd for C₂₇H₃₉N₃O₂438.30; found 439.

Preparation 12 Biphenyl-2-ylcarbamic Acid1-{9-[2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethylamino]nonyl}piperidin-4-ylEster

[0551] The product of Preparation 11 (196 mg, 0.43 mmol) was dissolvedin dichloroethane (4 mL) and sodium triacetoxyborohydride (101 mg, 0.48mmol) was added. The reaction mixture was stirred at ambient temperaturefor about 10 min. and then8-benzyloxy-5-(2,2-dihydroxyacetyl)-1H-quinolin-2-one (Preparation 6)(141 mg, 0.43 mmol) was added. LCMS analysis showed that the reactionwas completed after 2 h. Methanol (1 mL) was added to the reactionmixture and then sodium borohydride (18 mg, 0.48 mmol) was added slowly.After 1 hour, LCMS analysis showed that the reaction was completed. Thereaction mixture was then quenched with aqueous ammonium chloride andthis mixture was extracted with dichloromethane. The organic phase waswashed with saturated sodium bicarbonate (2×50 mL) and brine (10 mL).The organic phase was then dried over magnesium sulfate and concentratedto provide 315 mg of a yellow solid. The solid was purified by silicagel chromatography using 10% methanol in dichloromethane to afford thetitle compound (64 mg). MS m/z: [M+H⁺] calcd for C₄₃H₅₅N₄O₅ 730.40;found 731.

Example 2 Biphenyl-2-ylcarbamic Acid1-{9-[2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]nonyl}piperidin-4-ylEster Ditrifluoroacetate

[0552] A solution of the product of Preparation 12 (64 mg, 0.09 mmol) inmethanol (450 mL) was flushed with nitrogen. Palladium on carbon (10%,10 mg) was then added and the reaction mixture was placed under aballoon containing hydrogen and stirred. LCMS analysis showed that thereaction was completed after 9 h. The reaction mixture was then filteredand the filtrate was concentrated to provide a yellow crispy solid. Thesolid was purified by preparative HPLC (5-35 over 60 min) to afford thetitle compound (19 mg). MS m/z: [M+H⁺] calcd for C₃₈H₄₈N₄O₅ 641.36;found 641.

Preparation 138-Benzyloxy-5-[(R)-2-bromo-1-(tert-butyldimethylsilanyloxy)ethyl]-1H-quinolin-2-one

[0553] (a) 8-Benzyloxy-5-(2-Bromoacetyl)-1H-quinolin-2-one

[0554] 5-Acetyl-8-benzyloxy-1H-quinolin-2-one (Preparation 6) (20.0 g,68.2 mmol) was dissolved in dichloromethane (200 mL) and cooled to 0° C.Boron trifluoride diethyl etherate (10.4 mL, 82.0 mmol) was added viasyringe and the mixture was warmed to room temperature to give a thicksuspension. The suspension was heated at 45° C. (oil bath) and asolution of bromine (11.5 g, 72.0 mmol) in dichloromethane (100 mL) wasadded over 40 min. The mixture was kept at 45° C. for an additional 15min and then cooled to room temperature. The mixture was concentratedunder reduced pressure and then triturated with 10% aqueous sodiumcarbonate (200 mL) for 1 hour. The solids were collected on a Buchnerfunnel, washed with water (4×100 mL) and dried under reduced pressure.The product of two runs was combined for purification. The crude product(52 g) was triturated with 50% methanol in chloroform (500 mL) for 1hour. The product was collected on a Buchner funnel and washed with 50%methanol in chloroform (2×50 mL) and methanol (2×50 mL). The solid wasdried under reduced pressure to give the title compound (34.1 g) as apowder.

[0555] (b) 8-Benzyloxy-5-((R)-2-bromo-1-hydroxyethyl)-1H-quinolin-2-one

[0556] (R)-(+)-α,α-Diphenylprolinol (30.0 g, 117 mmol) andtrimethylboroxine (11.1 mL, 78 mmol) were combined in toluene (300 mL)and stirred at room temperature for 30 min. The mixture was placed in a150° C. oil bath and liquid was distilled off. Toluene was added in 20mL aliquots and distillation was continued for 4 h. A total of 300 mLtoluene was added. The mixture was then cooled to room temperature. A500 μL aliquot was evaporated to dryness and weighed (246 mg) todetermine that the concentration of catalyst was 1.8 M.

[0557] 8-Benzyloxy 5-(2-bromoacetyl)-1H-quinolin-2-one (90.0 g, 243mmol) was placed under nitrogen and tetrahydrofuran (900 mL) was addedfollowed by the catalyst described above (1.8 M in toluene, 15 mL, 27mmol). The suspension was cooled to −10±5° C. in an ice/isopropanolbath. Borane (1.0 M in THF, 294 mL, 294 mmol) was added over 4 h. Thereaction was then stirred an additional 45 min at −10° C. and thenmethanol (250 mL) was added slowly. The mixture was concentrated undervacuum and the residue was dissolved in boiling acetonitrile (1.3 L),filtered while hot and then cooled to room temperature. The crystalswere filtered, washed with acetonitrile and dried under vacuum to givethe title compound (72.5 g, 196 mmol, 81% yield, 95% ee, 95% pure byHPLC).

[0558] (c)8-Benzyloxy-5-[(R)-2-bromo-1-(tert-butyldimethylsilanyloxy)ethyl]-1H-quinolin-2-one

[0559] To the product of step (b) (70.2 g, 189 mmol) was addedN,N-dimethylformamide (260 mL) and this mixture was cooled in an icebath under nitrogen. 2,6-Lutidine (40.3 g, 376 mmol) was added over 5min and then tert-butyldimethylsilyl trifluoromethanesulfonate (99.8 g,378 mmol) was added slowly while maintaining the temperature below 20°C. The mixture was allowed to warm to room temperature for 45 min.Methanol (45 mL) was added to the mixture dropwise over 10 min and themixture was partitioned between ethyl acetate/cyclohexane(1:1, 500 mL)and water/brine (1:1, 500 mL). The organics were washed twice more withwater/brine (1:1, 500 mL each). The combined organics were evaporatedunder reduced pressure to give a light yellow oil. Two separate portionsof cyclohexane (400 mL) were added to the oil and distillation continueduntil a thick white slurry was formed. Cyclohexane (300 mL) was added tothe slurry and the resulting white crystals were filtered, washed withcyclohexane (300 mL) and dried under reduced pressure to give the titlecompound (75.4 g, 151 mmol, 80% yield, 98.6% ee).

Preparation 14 Biphenyl-2-ylcarbamic Acid1-{9-[(R)-2-(8-benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-(tert-butyldimethylsilanyloxy)ethylamino]nonyl}piperidin-4-ylEster

[0560] The product of Preparation 13 (3.9 g, 8.17 mmol) was added to asolution of the product of Preparation 11 (5.0 g, 11.4 mmol) in THF (20mL), followed by sodium bicarbonate (2.0 g, 24.5 mmol) and sodium iodide(1.8 g, 12.2 mmol). The reaction mixture was heated to 80° C. for 72 h.The reaction mixture was then cooled, diluted with dichloromethane (20mL) and the organic phase was washed with saturated sodium bicarbonate(2×50 mL) and brine (50 mL). The organic phase was then dried (magnesiumsulfate) and concentrated to give 6.5 g of a crude product. The crudeproduct was purified by chromatography on silica gel eluting with 3%methanol in dichloromethane to provide the title compound (1.4 g, 21%yield).

Preparation 15 Biphenyl-2-ylcarbamic Acid1-{9-[(R)-2-(8-benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethylamino]nonyl}piperidin-4-ylEster

[0561] Triethylamine hydrogen fluoride (376 μL, 2.3 mmol) was added to asolution of the product of Preparation 14 (1.3 g, 1.5 mmol) in THF (8mL) and the reaction mixture was stirred at ambient temperature. After 5h, the reaction was complete as determined by LCMS analysis. Thereaction mixture was then quenched with 1N NaOH until the pH was 14 andthen diluted with ethyl acetate (20 mL) and washed with 1N NaOH (20 mL)and brine (20 mL). The organic phase was then separated, dried overmagnesium sulfate, and concentrated to yield the title compound (1.1 g).

Example 3 Biphenyl-2-ylcarbamic Acid1-{9-[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydro-quinolin-5-yl)ethylamino]nonyl}piperidin-4-ylEster Ditrifluoroacetate

[0562] A solution of the product of Preparation 15 (1.1 g, 1.5 mmol) wasflushed with nitrogen and palladium on carbon (10%, 110 mg) was added.The reaction mixture was stirred under hydrogen at balloon pressure.Analysis by LCMS showed that the reaction was completed after 9 h. Thereaction mixture was then filtered and concentrated to yield a yellowsolid. The solid was purified by preparative HPLC (5-30 over 60 min) toafford the title compound (510 mg). MS m/z: [M+H⁺] calcd for C₃₈H₄₈N₄O₅641.36; found 641. HPLC method 10-70: 3.207. [α]²⁰ _(D)=−23.6 (c=1.0mg/mL, water).

Example 3A Biphenyl-2-ylcarbamic Acid1-{9-[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydro-quinolin-5-yl)ethylamino]nonyl}piperidin-4-ylEster Ditrifluoroacetate

[0563] Alternatively, the title compound was prepared as follows:

[0564] (a) 9-Bromononanal

[0565] To a 100-mL round-bottomed flask equipped with a magneticstirrer, addition funnel and temperature controller, under nitrogen, wasadded 9-bromononanol (8.92 g, 40 mmol) and dichloromethane (30 mL). Theresulting mixture was cooled to 5° C. and a solution of sodiumbicarbonate (0.47 g, 5.6 mmol) and potassium bromide (0.48 g, 4 mmol) inwater (10 mL) was added. 2,2,6,6-Tetramethyl-1-piperidinyloxy freeradical (TEMPO) (63 mg, 0.4 mmol) was added and then a 10 to 13% bleachsolution (27 mL) was added dropwise through the addition funnel at arate such that the temperature was maintained at about 8° C. (+/−2° C.)with an ice cold bath (over about 40 min.). After addition of the bleachwas complete, the mixture was stirred for 30 min. while maintaining thetemperature at about 0° C. A solution of sodium bisulfite (1.54 g) inwater (10 mL) was added and the resulting mixture was stirred at roomtemperature for 30 min. The layers of the mixture were then separated,and the milky aqueous layer was extracted with dichloromethane (1×20mL). The combined dichloromethane layers were then washed with water(1×30 mL), dried (MgSO₄), filtered and concentrated under reducedpressure to afford the title intermediate (8.3 g, 94% yield), which wasused without further purification in the next step.

[0566] (b) 9-Bromo-1,1-dimethoxynonane

[0567] To a 100 mL round-bottomed flask was added 9-bromononanal (7.2 g,32.5 mmol), methanol (30 mL) and trimethylorthoformate (4 mL, 36.5mmol). A solution of 4 N hydrochloric acid in dioxane (0.2 mL, 0.8 mmol)was added and the resulting mixture was refluxed for 3 h. The reactionmixture was then cooled to room temperature and solid sodium bicarbonate(100 mg, 1.2 mmol) was added. The resulting mixture was concentrated toone-fourth its original volume under reduced pressure and then ethylacetate (50 mL) was added. The organic layer was washed with water (2×40mL), dried (MgSO₄), filtered and concentrated under reduced pressure toafford the title intermediate (8.44 g, (97% yield)) as a liquid, whichas used in the next step without further purification.

[0568] (c) Biphenyl-2-ylcarbamic Acid1-(9,9-Dimethoxynonyl)piperidin-4-yl Ester

[0569] To a 50 mL three-necked, round-bottomed flask was addedbiphenyl-2-ylcarbamic acid piperidin-4-yl ester (1 g, 3.38 mmol) andacetonitrile (10 mL) to form a slurry. To this slurry was added9-bromo-1,1-dimethoxynonane (1.1 g, 1.3 mmol) and triethylamine (0.57 g,4.1 mmol) and the resulting mixture was heated at 65° C. for 6 h (thereaction was monitored by HPLC until starting material is <5%). Thereaction mixture was then cooled to room temperature at which time themixture formed a thick slurry. Water (5 mL) was added and the mixturewas filtered to collect the solid on a coarse fritted glass filer. Thesolid was washed with pre-mixed solution of acetonitrile (10 mL) andwater (5 mL) and then with another pre-mixed solution of acetonitrile(10 mL) and water (2 mL). The resulting solid was air dried to affordthe title intermediate (1.37 g, 84%, purity >96% by LC, 1H NMR) as awhite solid.

[0570] (d) Biphenyl-2-ylcarbamic Acid 1-(9-Oxononyl)piperidin-4-yl Ester

[0571] To a 500 mL round-bottomed flask with a magnetic stirrer wasadded biphenyl-2-ylcarbamic acid 1-(9,9-dimethoxynonyl)piperidin-4-ylester (7.7 g, 15.9 mmol) and then acetonitrile (70 mL) and aqueous 1Mhydrochloric acid (70 mL). The resulting mixture was stirred at roomtemperature for 1 h and then dichloromethane (200 mL) was added. Thismixture was stirred for 15 min. and then the layers were separated. Theorganic layer was dried (MgSO₄), filtered and concentrated under reducedpressure to afford the title intermediate (6.8 g), which was used in thenext step without further purification.

[0572] (e) Biphenyl-2-ylcarbamic Acid1-(9-{Benzyl-[(R)-2-(8-benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-(tert-butyldimethylsilanyloxy)ethyl]amino}nonyl)piperidin-4-ylEster

[0573] To a 300 mL round-bottomed flask was added5-[(R)-2-benzylamino-1-(tert-butyldimethylsilanyloxy)ethyl]-8-benzyloxy-1H-quinolin-2-one(5 g, 9.73 mmol), dichloromethane (100 mL) and glacial acetic acid (0.6mL, 10 mmol). This mixture was cooled to 0° C. using an ice bath andbiphenyl-2-ylcarbamic acid 1-(9-oxononyl)piperidin-4-yl ester (4.6 g,9.73 mmol) was added with stirring. This mixture was stirred at 0° C.for 30 minutes and then sodium triacetoxyborohydride (6.15 g, 29 mmol)was added in portions over 15 minutes. The reaction mixture was stirredat 0° C. to 10° C. for 2 hours and then aqueous saturated sodiumbicarbonate solution (50 mL) was added and this mixture was stirred for15 minutes. The layers were then separated and the organic layer waswashed with 5% aqueous sodium chloride solution (50 mL), dried (MgSO₄),filtered and concentrated under reduced pressure to afford the titleintermediate (8.5 g, 80% purity by HPLC), which was used without furtherpurification.

[0574] (f) Biphenyl-2-ylcarbamic Acid1-{9-[(R)-2-(tert-Butyldimethylsilanyloxy)-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]nonyl}piperidin-4-ylEster

[0575] To a 200 mL round-bottomed flask was added the intermediate fromStep E (8.5 g, 9 mmol), ethanol (100 mL) and glacial acetic acid (0.54mL, 18 mmol) and this mixture was stirred until the solid dissolved. Thereaction mixture was purged with hydrogen for 5 min. and then 10%palladium on carbon (1.7 g) was added. This mixture was stirred at roomtemperature while hydrogen was slowly bubbling through the reactionmixture until >95% conversion was observed by HPLC (about 8-9 h). Themixture was then filtered through a Celite pad and the solvent wasremoved under reduced pressure. The residue was purified by silica gelchromatography (15 g silica/1 g crude) using 5% MeOH in DCM/0.5% NH₄OH(10×150 mL), 8% MeOH in DCM/0.5% NH₄OH (10×150 mL) and 10% MeOH inDCM/0.5% NH₄OH (10×150 mL). The appropriate fractions were combined andthe solvent was removed under reduced pressure while maintaining thetemperature <35° C. to give the title intermediate (4.05 g, 97% purity).

[0576] (g) Biphenyl-2-ylcarbamic Acid1-{9-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]nonyl}piperidin-4-ylEster

[0577] To a 200 mL round-bottomed flask was added the intermediate fromStep F (4.05 g, 5.36 mmol) and dichloromethane (80 mL) and the resultingmixture was stirred until the solid dissolved. Triethylaminetrihydrofluoride (2.6 mL, 16 mmol) was added and stirring was continuedunder nitrogen for 18 to 20 h. Methanol (20 mL) was added and thensaturated aqueous sodium bicarbonate (50 mL) was added slowly and themixture was stirred for 15 min. The layers were then separated and theorganic layer was washed with saturated aqueous sodium chloride solution(20 mL), dried (MgSO₄), filtered and concentrated under reduced pressureto afford the title compound (3.5 g, 98% purity by HPLC) as a yellowsolid.

Example 3B Biphenyl-2-ylcarbamic Acid1-{9-[(R)-2-Hydroxy-2-(8-Hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]nonyl}piperidin-4-ylEster Naphthalene-1,5-disulfonic Acid Salt

[0578] Biphenyl-2-ylcarbamic acid1-{9-[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]nonyl}piperidin-4-ylester (1.0 g, 1.56 mmol, free base) was dissolved in methanol (10 mL;low water content). A solution of naphthalene-1,5-disulfonic acid (0.45g, 1.56 mmol) in methanol (5 mL; low water content) was added and thereaction mixture was stirred at 30° C. for two hours and then at roomtemperature overnight (18 h). The resulting thick slurry was filteredand the filtrate cake was washed with methanol (5 mL) and then dried togive the title compound (1.16 g, 80% yield) as off-white crystallinesolid.

Preparation 16N-{2-Benzyloxy-5-[(R)-2-bromo-1-(tert-butyldimethylsilanyloxy)ethyl]phenyl}-formamide

[0579] (R)-2-Bromo-1-(3-formamido-4-benzyloxyphenyl)ethanol (9.9 g, 28mmol) was dissolved in dimethylformamide (36 mL). Imidazole (2.3 g, 34mmol) and tert-butyldimethylsilyl chloride (4.7 g, 31 mmol) were added.The solution was stirred under nitrogen atmosphere for 72 h. Additionalimidazole (0.39 g, 5.7 mmol) and tert-butyldimethylsilyl chloride (0.64g, 4.3 mmol) were added and the reaction was stirred for an additional20 h. The reaction mixture was then diluted with a mixture of isopropylacetate (53 mL) and hexanes (27 mL) and transferred to a separatoryfunnel. The organic layer was washed twice with a mixture of water (27mL) and saturated aqueous sodium chloride (27 mL) followed by a finalwash with saturated aqueous sodium chloride (27 mL). The organic layerwas dried over sodium sulfate. Silica gel (23.6 g) and hexanes (27 mL)were added and the suspension was stirred for 10 min. The solids wereremoved by filtration and the filtrate concentrated under vacuum. Theresidue was crystallized from hexanes (45 mL) to afford 8.85 g (19 mmol,68%) of the title compound as a solid. MS m/z: [M+H⁺] calcd forC₂₂H₃₀NO₃SiBr 464.1; found 464.2.

[0580] The starting material,(R)-2-bromo-1-(3-formamido-4-benzyloxyphenyl)ethanol, can be prepared asdescribed in U.S. Pat. No. 6,268,533 B1; or R. Hett et al., OrganicProcess Research and Development, 1998, 2:96-99; or using proceduressimilar to those described in Hong et al., Tetrahedron Lett., 1994,35:6631; or similar to those described in U.S. Pat. No. 5,495,054.

Preparation 17 Biphenyl-2-ylcarbamic Acid1-{9-[(R)-2-(4-Benzyloxy-3-formylaminophenyl)-2-(tert-butyldimethylsilanyloxy)ethylamino]nonyl}piperidin-4-ylEster

[0581] The product of Preparation 16 (500 mg, 1.008 mmol) and sodiumiodide (243 mg, 1.62 mmol) were stirred in tetrahydrofuran (0.5 mL) for15 min at ambient temperature. The product of Preparation 11, (564 mg,1.29 mmol) and sodium bicarbonate (272 mg, 3.24 mmol) were then addedand the reaction mixture was heated at 80° C. for 24 h. The reactionmixture was then allowed to cool. Water (2 mL) was then added and themixture was extracted with dichloromethane (2×2 mL). The combinedorganic extracts were washed with 1M hydrochloric acid (2×1 mL), dried(magnesium sulfate) and the solvent was removed under reduced pressure.The crude residue was purified by flash chromatography (5-10%methanol/dichloromethane) to give the title compound (360 mg, 0.44 mmol,41% yield). HPLC (10-70) R_(t)=4.96; MS m/z: [M+H⁺] calcd for C₄₉H₆₈N₄O₅821.51; found 821.9.

Preparation 18 Biphenyl-2-ylcarbamic Acid1-{9-[(R)-2-(4-Benzyloxy-3-formylaminophenyl)-2-hydroxyethylamino]nonyl}piperidin-4-ylEster

[0582] To a stirred solution of the product of Preparation 17 (360 mg,0.44 mmol) in tetrahydrofuran (2.2 mL) at ambient temperature was addedtriethylamine trihydrofluoride (108 μL, 0.66 mmol). The reaction mixturewas stirred for 24 h and then diluted with dichloromethane (5 mL) andwashed with 1 M hydrochloric acid (2 mL) and saturated aqueous sodiumbicarbonate (2 mL). The organic phase was dried (magnesium sulfate) andthe solvent was removed under reduced pressure. The crude title compoundwas used directly in the next step without further purification. HPLC(10-70) R_(t)=4.6; MS m/z: [M+H⁺] calcd for C₄₃H₅₄N₄O₅ 707.43; found707.8.

Example 4 Biphenyl-2-ylcarbamic Acid1-{9-[(R)-2-(3-Formylamino-4-hydroxyphenyl)-2-hydroxyethylamino]nonyl}piperidin-4-ylEster Ditrifluoroacetate

[0583] Palladium (10 wt. % (dry basis) on activated carbon) (124 mg) wasadded to a stirred solution of the product of Preparation 18 (311 mg,0.44 mmol) in ethanol (4 mL) and the reaction mixture was placed underan atmosphere of hydrogen. After stirring for 12 h, the reaction mixturewas filtered through a pad of Celite, washed with methanol (2 mL) andthe solvent was removed under reduced pressure. The resulting residuewas purified by preparative HPLC to give the title compound (41 mg).HPLC (10-70) R_(t)=3.0; MS m/z: [M+H⁺] calcd for C₃₆H₄₈N₄O₅ 617.39;found 617.5.

Example 5 Biphenyl-2-ylcarbamic Acid1-{9-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2,3,4-tetrahydroquinolin-5-yl)ethylamino]nonyl}piperidin-4-ylEster Ditrifluoroacetate

[0584] Palladium (10 wt. % (dry basis) on activated carbon) (80 mg) wasadded to a stirred solution of the product of Example 3 (80 mg, 0.11mmol) in ethanol (1.1 mL) and the reaction mixture was placed under anatmosphere of hydrogen. The reaction mixture was stirred for 12 h, andthen filtered through a pad of Celite, washed with methanol (2 mL) andthe solvent removed under reduced pressure. The crude material wasresubjected to the above conditions to ensure complete reaction. Theresulting residue was purified by preparative HPLC to yield the titlecompound (6 mg). HPLC (10-70) R_(t)=3.23; MS m/z: [M+H⁺] calcd forC₃₈H₅₀N₄O₅ 643.39; found 643.7.

Preparation 19 3-[4-(Biphenyl-2-ylcarbamoyloxy)piperidin-1-yl]propionicAcid Methyl Ester

[0585] Methyl 3-bromopropionate (553 μL, 5.07 mmol) was added to astirred solution of the product of Preparation 8 (1.00 g, 3.38 mmol) andDIPEA (1.76 mL, 10.1 mmol) in acetonitrile (34 mL) at 50° C. and thereaction mixture was heated at 50° C. overnight. The solvent was thenremoved under reduced pressure and the residue was dissolved indichloromethane (30 mL). The resulting solution was washed withsaturated aqueous sodium bicarbonate solution (10 mL), dried (magnesiumsulfate) and the solvent was removed under reduced pressure. The cruderesidue was purified by column chromatography (5-10% MeOH/DCM) to givethe title compound (905 mg, 70%).

Preparation 20 3-[4-(Biphenyl-2-ylcarbamoyloxy)piperidin-1-yl]propionicAcid

[0586] A stirred solution of the product of Preparation 19 (902 mg, 2.37mmol) and lithium hydroxide (171 mg, 7.11 mmol) in 50% THF/H₂O (24 mL)was heated at 30° C. overnight, and then acidified with concentratedhydrochloric acid and lyophilized to give the title compound (˜100%yield, also contains LiCl salts).

Preparation 21{5-[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-(tert-butyldimethyl-silanyloxy)ethylamino]pentyl}carbamicAcid tert-Butyl Ester

[0587] The product of Preparation 13 (600 mg, 1.23 mmol) andN-tert-butoxycarbonyl-1,5-diaminopentane (622 mg, 3.07 mmol) weredissolved in dimethyl sulfoxide (1.23 mL) and heated to 105° C. for 6 h.The reaction mixture was then cooled and diluted with ethyl acetate (10mL) and washed with saturated aqueous sodium bicarbonate solution (4mL). The organic phase was dried (magnesium sulfate) and the solvent wasremoved under reduced pressure. The crude residue was purified by columnchromatography (5-10% methanol/dichloromethane) to give the titlecompound (˜100% yield).

Preparation 225-[(R)-2-(5-Aminopentylamino)-1-(tert-butyldimethylsilanyloxy)ethyl]-8-benzyloxy-1H-quinolin-2-one

[0588] A solution of the product of Preparation 21 (800 mg, 1.31 mmol)in trifluoroacetic acid/dichloromethane (25%, 12 mL) was stirred atambient temperature for 1 hour. The solvent was then removed underreduced pressure and the crude residue was dissolved in dichloromethane(15 mL) and washed with 1N sodium hydroxide (8 mL). The organic phasewas separated, dried (magnesium sulfate) and the solvent was removedunder reduced pressure to give the title compound (509 mg, 81% yieldover 2 steps).

Preparation 23 Biphenyl-2-ylcarbamic Acid1-(2-{5-[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-(tert-butyldimethylsilanyloxy)ethylamino]pentylcarbamoyl}ethyl)piperidin-4-ylEster

[0589] To the product of Preparation 20 (417 mg, 1.13 mmol) and HATU(430 mg, 1.13 mmol) was added the product of Preparation 22 (458 mg,0.90 mmol) in DMF (1.8 mL), followed by DIPEA (204 μL, 1.17 mmol). Thereaction mixture was stirred at 50° C. for 12 h, and then the solventwas removed under reduced pressure. The crude residue was dissolved indichloromethane (10 mL). The resulting solution was washed withsaturated aqueous sodium bicarbonate solution (4 mL), dried (magnesiumsulfate) and the solvent was removed under reduced pressure. The cruderesidue was purified by column chromatography (5-10%methanol/dichloromethane and 0.5% NH₄OH) to give the title compound (240mg, 3 1% yield).

Preparation 24 Biphenyl-2-ylcarbamic Acid1-(2-{5-[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethylamino]pentylcarbamoyl}ethyl)piperidin-4-ylEster

[0590] To a stirred solution of the product of Preparation 23 (240 mg,0.28 mmol) in dichloromethane (2.8 mL) was added triethylaminetrihydrofluoride (91 μL, 0.56 mmol). The reaction mixture was stirredfor 10 h, and then diluted with dichloromethane (10 mL). The resultingsolution was then washed with saturated aqueous sodium bicarbonatesolution (5 mL), and then the organic phase was dried (magnesiumsulfate) and the solvent was removed under reduced pressure to give thetitle compound (209 mg, 100% yield).

Example 6 Biphenyl-2-ylcarbamic Acid1-(2-{5-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]pentylcarbamoyl}ethyl)piperidin-4-ylEster, Ditrifluoroacetate

[0591] To a stirred solution of the product of Preparation 24 (209 mg,0.28 mmol) in ethanol (2.8 mL) was added palladium (10 wt. % (dry basis)on activated carbon) (81 mg) and the reaction mixture was placed underan atmosphere of hydrogen and stirred overnight. The reaction mixturewas then filtered and solvent was removed under reduced pressure. Thecrude residue was purified by preparative HPLC to give the titlecompound (58 mg). HPLC (10-70) R_(t)=2.30; MS m/z: [M+H⁺] calcd forC₃₇H₄₅N₅O₆ 656.34; found 656.6; [α]²⁰ _(D)=−6.5 (c=1.0 mg/mL, water).

Example 6A Biphenyl-2-ylcarbamic Acid1-(2-{5-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]pentylcarbamoyl}ethyl)piperidin-4-ylEster

[0592] Alternatively, the title compound can be prepared as follows:

[0593] (a) 5-Chloropentanal

[0594] To a 2 L three-necked round-bottomed flask, equipped with amagnetic stirrer, addition funnel and temperature controller undernitrogen, was added 5-chloropentanol (53 g, 0.433 mol) anddichloromethane (300 mL). This mixture was cooled to 5° C. and asolution of sodium bicarbonate (5 g, 0.059 mol) and potassium bromide(5.1 g, 0.043 mol) in water (225 mL) was added.2,2,6,6-Tetramethyl-1-piperidinyloxy free radical (TEMPO) (63 mg, 0.4mmol) was added and then a 10 to 13% bleach solution (275 mL) was addeddropwise through the addition funnel at a rate such that the temperaturewas maintained at about 8° C. (+/−2° C.) with an ice cold bath (overabout 45 min.). After addition of the bleach was complete, the mixturewas stirred for 30 min. while maintaining the temperature at about 5° C.A solution of sodium bisulfite (4 g) in water (30 mL) was added and theresulting mixture was stirred at room temperature for 30 min. The layersof the mixture were then separated, and the aqueous layer was extractedwith dichloromethane (1×50 mL). The combined dichloromethane layers werethen washed with water (1×50 mL), dried (MgSO₄), filtered andconcentrated under reduced pressure to afford the title compound (53 g).The product was distilled at 65° C./8 torr to afford the title compound(31.16 g) as an orange oil (GC purity was 70 to 80%).

[0595] The product was further purified by adding the crude material (4g) to a mixture of ethanol (920 mL), ethyl acetate (12 mL) and water (4mL). Sodium bisulfite (4 g) was added and the mixture was heated toreflux for 4 h and then cooled to room temperature and stirred for 14 hat room temperature to form a very thick slurry. The solids werefiltered on a coarse fritted filter, washed with the solvent mixture (5mL) and the solids were dried on the filter to afford 8.4 g of thebisulfite adduct. This material was then added to MTBE (20 mL) andaqueous 1 N sodium hydroxide (45 mL) was added with vigorous stirring.The resulting biphasic mixture was stirred vigorously until all thesolids had dissolved (about 15 min) and then the layers were separated.The aqueous layer was extracted with MTBE (20 mL) and the combined MTBElayers were dried (MgSO₄), filtered and concentrated to afford 3.46 g ofthe title compound as a colorless liquid (GC purity >90%).

[0596] (b)5-[(R)-2-[Benzyl-(5-chloropentyl)amino]-1-(tert-butyldimethylsilanyloxy)ethyl]-8-benzyloxy-1H-quinolin-2-one

[0597] To a 1 L three-necked round-bottomed flask was added the productof Preparation 28 (48.4 g, 94 mmol), dichloromethane (400 mL) andglacial acetic acid (11.3 mL). This mixture was stirred at 0° C. (icebath) and the product from step (a) (12.5 g, 103.6 mmol) was added andstirring was continued for 15 min. Sodium triacetoxyborohydride (59.8 g,282 mmol) was then added in portions over a 15 min. period and theresulting mixture was stirred at 0° C. to 10° C. for 2 h. Aqueoussaturated sodium bicarbonate solution (200 mL) was then added slowly(gas evolution) and stirring was continued for 15 min. The pH of thesolution was then adjusted with solid sodium carbonate to a pH of about9 and the layers were separated. The organic layer was washed withaqueous 5% sodium chloride solution (200 mL), dried (MgSO₄), filteredand concentrated under reduced pressure to afford the title compound (53g).

[0598] (c)5-[(R)-2-[(5-N,N-Diformylaminopentyl)benzylamino]-1-(tert-butyldimethyl-silanyloxy)ethyl]-8-benzyloxy-1H-quinolin-2-one

[0599] To a stirred solution of the product of step (b) (26.5 g, 42.8mmol) in 1-methyl-2-pyrrolidinone (175 mL) was added sodiumdiformylamide (6.1 g, 64.2 mmol) and sodium iodide (2.13 g, 14.3 mmol).The reaction flask was flushed with nitrogen and then the mixture washeated at 65° C. for 8 h. The mixture was then cooled to roomtemperature and water (300 mL) and ethyl acetate (100 mL) were added.This mixture was stirred for 10 min. and then the layers were separated.The aqueous layer was extracted with ethyl acetate (150 mL) and thecombined organic layers were washed with water (300 mL), aqueous 50%brine solution (300 mL), water (300 mL), dried (MgSO₄) filtered andconcentrate to afford the title compound (23.3 g).

[0600] (d)5-[(R)-2-[(5-Aminopentyl)benzylamino]-1-(tert-butyldimethylsilanyloxy)ethyl]-8-benzyloxy-1H-quinolin-2-one

[0601] To a stirred solution of the product from step (c) (10.5 g, 16mmol) in methanol (75 mL) was added p-toluenesulfonic acid (7.42 g. 39mmol). The resulting mixture was heated at 40° C. for 15 h and thenconcentrated under reduced pressure to about half its volume. Methanol(70 mL) was added and the mixture was heated at 50° C. for 2 h and thenconcentrated under reduced pressure. Water (100 mL), methanol (50 mL)and MTBE (100 mL) were added and this mixture was stirred for 15 min andthen the layers were separated. To the aqueous layer was added aqueous 1N sodium hydroxide (45 mL) and MTBE (100 mL), and this mixture wasstirred for 15 min. The layers were then separated and the aqueous layerwas extracted with MTBE (100 mL). The combined MTBE layers were dried(MgSO₄), filtered and concentrated to afford the title compound as ayellow oil (7.3 g). This material contained about 13% (by HPLC) of thecorresponding des-tert-butyldimethylsilyl compound.

[0602] (e) 3-[4-(Biphenyl-2-ylcarbamoyloxy)piperidin-1-yl]propionic Acid

[0603] To a solution of the product of Preparation 8 (50 g, 67.6 mmol)in dichloromethane (500 mL) was added acrylic acid (15.05 mL, 100 mmol).The resulting mixture was heated at 50° C. under reflux for 18 h andthen the solvent was removed. Methanol (600 mL) was added and thismixture was heated at 75° C. for 2 h and then cooled to room temperatureto form a thick slurry. The solid was collected by filtration, washedwith methanol (50 mL) and air dried to afford the title compound (61g, >96% purity) as white powder.

[0604] (f) Biphenyl-2-ylcarbamic Acid1-[2-(5-{Benzyl-[(R)-2-(8-benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-(tert-butyldimethylsilanyloxy)ethyl]amino}-pentylcarbamoyl)ethyl]piperidin-4-ylEster

[0605] A mixture of the product of step (e) (3.68 g, 10 mmol) andN,N-dimethylformamide (50 mL) was heated at 60° C. until the solidcompletely dissolved and then cooled to room temperature. The product ofstep (d) (6 g, 10 mmol) and diisopropylethylamine (3.5 mL) was added andthe reaction mixture was cooled to 0° C. PyBOP (6.25 g, 12 mmol) wasadded in one portion and the reaction mixture was stirred at 0° C. toroom temperature for 2 hours. The reaction mixture was then poured intocold water (500 mL) with stirring and the pH of the resulting mixturewas adjusted to about 2 using aqueous 1 M hydrochloric acid. Thismixture was stirred for 15 min and then filtered to collect the solid,which was washed with water (100 mL) and dried to afford the titlecompound (8.7 g, HPLC purity >95%) as an off-white solid.

[0606] (g) Biphenyl-2-ylcarbamic Acid1-(2-{5-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]pentylcarbamoyl}ethyl)piperidin-4-ylEster

[0607] The product of step (f) can be deprotected using essentially thesame procedures as those described in Preparation 24 and Example 6 toafford the title compound.

Preparation 25 2-(N-Benzyloxycarbonyl-N-methylamino)ethanal

[0608] (a) 2-(N-Benzyloxycarbonyl-N-methylamino)ethanol

[0609] Benzyl chloroformate (19 g, 111.1 mmol) in THF (20 mL) was addeddropwise over 15 min to a stirred solution of 2-(methylamino)ethanol (10g, 133.3 mmol) in THF (100 mL) and aqueous sodium carbonate (100 mL) at0° C. The reaction mixture was stirred at 0° C. for 12 h and thenextracted with EtOAc (2×200 mL). The organic layer was washed withaqueous sodium carbonate (200 mL) and dried (potassium carbonate) andsolvent was removed under reduced pressure to give the title compound(22.5 g, 97% yield).

[0610] (b) 2-(N-Benzyloxycarbonyl-N-methylamino)ethanal

[0611] DMSO (71 mL, 1 mol) and DIPEA (87.1 mL, 0.5 mol) were added to astirred solution of the product of step (a) (20.9 g, 0.1 mol) indichloromethane (200 mL) at −10° C. The reaction mixture was stirred at−10° C. for 15 min and then sulfur trioxide pyridine complex (79.6 g,0.5 mol) was added and the resulting mixture was stirred for 1 hour. Thereaction mixture was quenched with addition of 1M hydrochloric acid (200mL). The organic layer was separated and washed with saturated aqueoussodium bicarbonate (100 mL), brine (100 mL), dried (potassium carbonate)and solvent removed under reduced pressure to give the title compound(20.7 g, ˜100% yield).

Preparation 26 Biphenyl-2-ylcarbamic Acid1-[2-(methylamino)ethyl]piperidin-4-yl Ester

[0612] To a stirred solution of the product of Preparation 25 (20.7 g,100 mmol) and the product of Preparation 8 (25 g, 84.7 mmol) in MeOH(200 mL) was added sodium triacetoxyborohydride (21.2 g, 100 mmol). Thereaction mixture was stirred for 12 h at ambient temperature and then itwas quenched with 2M hydrochloric acid and the solvent was removed underreduced pressure. The residue was dissolved in ethyl acetate (200 mL)and washed with saturated aqueous sodium bicarbonate solution (100 mL)and brine (50 mL), and then dried (magnesium sulfate) and the solventwas removed under reduced pressure. The crude residue was purified bycolumn chromatography (50-90% EtOAc/hexanes) to givebiphenyl-2-ylcarbamic acid1-[2-(benzyloxycarbonyl-methylamino)ethyl]piperidin-4-yl ester as anoil.

[0613] The oil was dissolved in methanol (100 mL) and palladium (10 wt.% (dry basis) on activated carbon) (5 g) was added. The reaction mixturewas stirred under hydrogen (30 psi) for 12 h and then filtered throughCelite, which was washed with methanol, and solvent was evaporated togive the title compound (13.2 g, 44% yield).

Preparation 27 Biphenyl-2-ylcarbamic Acid1-{2-[(6-Bromohexanoyl)methylamino]ethyl}piperidin-4-yl Ester

[0614] 6-Bromohexanoyl chloride (3.23 mL, 21.1 mmol) was added to astirred solution of the product of Preparation 26 (6.2 g, 17.6 mmol) andDIPEA (6.13 mL, 35.2 mmol) in dichloroethane (170 mL). The reactionmixture was stirred for 1 hour and it was then diluted with EtOAc (250mL) and washed with saturated aqueous sodium bicarbonate solution (2×200mL) and brine (200 mL), and then dried (magnesium sulfate). The solventwas removed under reduced pressure to give the title compound (6.6 g,73% yield).

Preparation 288-Benzyloxy-5-[(R)-2-(N-benzylamino)-1-(tert-butyldimethylsilanyloxy)ethyl]-1H-quinolin-2-one

[0615] A stirred solution of the product of Preparation 13 (1.00 g, 2.05mmol) and benzylamine (493 μL, 4.51 mmol) in DMSO (1.7 mL) was heated at105° C. for 4 h. The reaction mixture was allowed to cool and was thendiluted with EtOAc (10 mL) and the organic layer was washed withsaturated aqueous ammonium chloride solution (5 mL) and 1N sodiumhydroxide (5 mL), dried (MgSO₄) and solvent removed under reducedpressure. The crude residue was purified by column chromatography (50%EtOAc/hexanes) to give the title compound (700 mg, 67%). MS m/z: [M+H⁺]calcd for C₃₁H₃₈N₂O₃Si 515.27; found 515.5.

Preparation 29 Biphenyl-2-ylcarbamic Acid1-{2-[(6-{Benzyl-[(R)-2-(8-benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-(tert-butyldimethylsilanyloxy)ethyl]amino}hexanoyl)-methylamino]ethyl}piperidin-4-ylEster

[0616] To a stirred solution of the product of Preparation 28 (807 mg,1.57 mmol) and DIPEA (819 μL, 4.7 mmol) in acetonitrile (3.14 mL) wasadded the product of Preparation 27 (995 mg, 1.88 mmol). The reactionmixture was heated to 80° C. for 24 h. The solvent was removed underreduced pressure and the residue was dissolved in EtOAc (10 mL) and thenwashed with saturated aqueous sodium bicarbonate solution (5 mL), dried(magnesium sulfate), and the solvent removed under reduced pressure. Thecrude material was purified by column chromatography (4-6% MeOH/DCM) toobtain the title compound (452 mg, 30% yield).

Preparation 30 Biphenyl-2-ylcarbamic Acid1-{2-[(6-{Benzyl-[(R)-2-(8-benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethyl]amino}hexanoyl)methylamino]ethyl}piperidin4-yl Ester

[0617] To a stirred solution of the product of Preparation 29 (452 mg,0.47 mmol) in dichloromethane (4.7 mL) was added triethylaminetrihydrofluoride (116 μL, 0.71 mmol). The reaction mixture was stirredfor 10 h. and then it was diluted with dichloromethane (10 mL) andwashed with saturated aqueous sodium bicarbonate solution (5 mL). Theorganic phase was then dried (MgSO₄) and the solvent was removed underreduced pressure to give the title compound (100% yield).

Example 7 Biphenyl-2-ylcarbamic Acid1-[2-({6-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]hexanoyl}methylamino)ethyl]piperidin-4-ylEster Ditrifluoroacetate

[0618] To a stirred solution of the product of Preparation 30 (400 mg,0.47 mmol) in ethanol (4.7 mL) was added palladium (10 wt. % (dry basis)on activated carbon) (160 mg) and the reaction mixture was placed underan atmosphere of hydrogen and stirred overnight. The reaction mixturewas then filtered and solvent was removed under reduced pressure. Thecrude residue was purified by preparative HPLC to give the titlecompound (73 mg). HPLC (10-70) R_(t)=2.33; MS m/z: [M+H⁺] calcd forC₃₈H₄₇N₅O₆ 670.36; found 670. [α]²⁰ _(D)=−9.4 (c=1.0 mg/mL, water).

Preparation 31 Biphenyl-2-ylcarbamic Acid1-[2-(4-(Aminomethyl)phenylcarbamoyl)-ethyl]piperidin-4-yl Ester

[0619] To a stirred solution of4-(N-tert-butoxycarbonylaminomethyl)aniline (756 mg, 3.4 mmol), theproduct of Preparation 20 (1.5 g, 4.08 mmol) and HATU (1.55 g, 4.08mmol) in DMF (6.8 mL) was added DIPEA (770 μL, 4.42 mmol). The reactionmixture was stirred at 50° C. overnight and then the solvent was removedunder reduced pressure. The resulting residue was dissolved indichloromethane (20 mL) and washed with saturated aqueous sodiumbicarbonate solution (10 mL). The organic phase was then dried(magnesium sulfate) and the solvent was removed under reduced pressure.The crude product was purified by flash chromatography (5-10% MeOH/DCM)to give a solid, which was dissolved in TFA/DCM (25%, 30 mL) and stirredat room temperature for 2 h. The solvent was then removed under reducedpressure and the crude residue was dissolved in dichloromethane (30 mL)and washed with 1N sodium hydroxide (15 mL). The organic phase wasseparated, dried (magnesium sulfate) and the solvent was removed underreduced pressure to give the title compound (1.5 g, 94% over 2 steps).

Preparation 32 Biphenyl-2-ylcarbamic Acid1-[2-(4-{[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-(tert-butyldimethylsilanyloxy)ethylamino]methyl}phenylcarbamoyl)ethyl]-piperidin-4-ylEster

[0620] A solution of the product of Preparation 31 (489 mg, 1.04 mmol),the product of Preparation 13 (610 mg, 1.25 mmol), sodium bicarbonate(262 mg, 3.12 mmol) and sodium iodide (203 mg, 1.35 mmol) in THF (0.52mL) were heated at 80° C. for 12 h. The reaction mixture was dilutedwith dichloromethane (10 mL) and washed with saturated aqueous sodiumbicarbonate solution (5 mL). The organic phase was dried (MgSO₄) and thesolvent was removed under reduced pressure. The crude residue waspurified by flash chromatography (10% MeOH/DCM) to give the titlecompound as a solid (687 mg, 77% yield).

Preparation 33 Biphenyl-2-ylcarbamic Acid1-[2-(4-{[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethylamino]methyl}phenylcarbamoyl)ethyl]piperidin-4-ylEster

[0621] To a stirred solution of the product of Preparation 32 (687 mg,0.8 mmol) in dichloromethane (8 mL) was added triethylaminetrihydrofluoride (261 μL, 1.6 mmol). The reaction mixture was stirredfor 10 h and then was diluted with dichloromethane (20 mL) and washedwith saturated aqueous sodium bicarbonate solution (10 mL). The organicphase was then dried (magnesium sulfate) and the solvent was removedunder reduced pressure to yield the title compound (500 mg, 81% yield).

Example 8 Biphenyl-2-ylcarbamic Acid1-[2-(4-{[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenylcarbamoyl)ethyl]piperidin-4-ylEster Ditrifluoroacetate

[0622] To a stirred solution of the product of Preparation 33 (500 mg,0.65 mmol) in ethanol (6.5 mL) was added palladium (10 wt. % (dry basis)on activated carbon) (200 mg) and the reaction mixture was placed undera hydrogen atmosphere and stirred overnight. The reaction mixture wasthen filtered and the solvent was removed under reduced pressure. Thecrude residue was purified by preparative HPLC to give the titlecompound (81 mg, 2 TFA salt). HPLC (10-70) R_(t)=2.41; MS m/z: [M+H⁺]calcd for C₃₉H₄₁N₅O₆ 676.32; found 676.5.

Preparation 34 Biphenyl-2-ylcarbamic Acid1-(2-tert-Butoxycarbonylaminoethyl)piperidin-4-yl Ester

[0623] To a stirred solution of the product of Preparation 8 (2.00 g,6.76 mmol) and DIPEA (3.54 mL, 20.3 mmol) in acetonitrile (67.6 mL) at50° C. was added 2-tert-butoxycarbonylaminoethyl bromide (1.82 g, 8.11mmol) and the reaction mixture was heated at 50° C. overnight. Thesolvent was then removed under reduced pressure and the residue wasdissolved in dichloromethane (60 mL) and washed with saturated aqueoussodium bicarbonate solution (30 mL). The organic phase was dried(magnesium sulfate) and the solvent was removed under reduced pressure.The crude residue was purified by column chromatography (5% MeOH/DCM) toyield the title compound as a solid (2.32 g, 78% yield).

Preparation 35 Biphenyl-2-ylcarbamic Acid 1-(2-Aminoethyl)piperidin-4-ylEster

[0624] The product of Preparation 34 was dissolved in TFA/DCM (25%, 52mL) and stirred at room temperature for 2 h. The solvent was thenremoved under reduced pressure and the crude residue dissolved indichloromethane (30 mL) and washed with 1N sodium hydroxide (15 mL). Theorganic phase was separated, dried (magnesium sulfate) and the solventwas removed under reduced pressure to give the title compound (1.61 g,90% yield).

Preparation 36 Biphenyl-2-ylcarbamic Acid1-[2-(4-Aminomethylbenzoylamino)ethyl]piperidin-4-yl Ester

[0625] To a stirred solution of the product of Preparation 35 (339 mg, 1mmol), 4-(tert-butoxycarbonylaminomethyl)benzoic acid (301 mg, 1.2 mmol)and HATU (456 mg, 1.2 mmol) in DMF (2 mL) was added DIPEA (226 μL, 1.3mmol). The reaction mixture was stirred at room temperature overnightand then the solvent was removed under reduced pressure. The resultingresidue was dissolved in dichloromethane (20 mL) and washed withsaturated aqueous sodium bicarbonate solution (10 mL). The organic phasewas dried (magnesium sulfate) and the solvent was removed under reducedpressure. The crude product was dissolved in TFA/DCM (25%, 10 mL) andthis mixture was stirred at room temperature for 2 h. The solvent wasremoved under reduced pressure and the crude residue was dissolved indichloromethane (15 mL) and washed with 1N sodium hydroxide (5 mL). Theorganic phase was separated, dried (magnesium sulfate) and the solventwas removed under reduced pressure to afford the title compound (472 mg,˜100% over 2 steps).

Preparation 37 Biphenyl-2-ylcarbamic Acid1-[2-(4-{[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-(tert-butyldimethylsilanyloxy)ethylamino]methyl}benzoylamino)ethyl]-piperidin-4-ylEster

[0626] A solution of the product of Preparation 36 (520 mg, 1.1 mmol),the product of Preparation 13 (634 mg, 1.3 mmol), sodium bicarbonate(277 mg, 3.3 mmol) and sodium iodide (215 mg, 1.43 mmol) in THF (0.55mL) was heated at 80° C. for 12 h. The reaction mixture was then dilutedwith dichloromethane (10 mL) and washed with saturated aqueous sodiumbicarbonate solution (5 mL). The organic phase was then dried (magnesiumsulfate) and the solvent was removed under reduced pressure. The cruderesidue was purified by flash chromatography (5-10% MeOH/DCM) to givethe title compound as a solid (316 mg, 33% yield).

Preparation 38 Biphenyl-2-ylcarbamic Acid1-[2-(4-{[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethylamino]methyl}benzoylamino)ethyl]piperidin-4-ylEster

[0627] To a stirred solution of the product of Preparation 37 (316 mg,0.36 mmol) in dichloromethane (3.6 mL) was added triethylaminetrihydrofluoride (117 μL, 0.72 mmol). The reaction mixture was stirredfor 10 h and was then diluted with dichloromethane (10 mL) and washedwith saturated aqueous sodium bicarbonate solution (5 mL). The organicphase was dried (MgSO₄) and the solvent was removed under reducedpressure to give the title compound, which was used directly in the nextstep (100% yield).

Example 9 Biphenyl-2-ylcarbamic Acid1-[2-(4-{[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}benzoylamino)ethyl]piperidin-4-ylEster Ditrifluoroacetate

[0628] To a stirred solution of the product of Preparation 38 (275 mg,0.36 mmol) in ethanol (3.6 mL) was added palladium (10 wt. % (dry basis)on activated carbon) (275 mg) and the reaction mixture was placed undera hydrogen atmosphere and stirred overnight. The reaction mixture wasthen filtered and the solvent was removed under reduced pressure. Thecrude residue was purified by preparative HPLC to yield the titlecompound (6 mg, 2 TFA salt). HPLC (10-70) R_(t)=2.26; MS m/z: [M+H⁺]calcd for C₃₉H₄₁N₅O₆ 676.32; found 676.5.

Preparation 39 Biphenyl-2-ylcarbamic Acid 1-(2-Aminoethyl)piperidin-4-ylEster

[0629] 2-tert-Butoxycarbonylaminoethyl bromide (1.22 g, 5.44 mmol) wasadded to a solution of the product of Preparation 8 (1.46 g, 4.95 mmol)and diisopropylethylamine (1.03 mL, 5.94 mmol) in acetonitrile (24 mL).The reaction mixture was stirred at 65° C. for 12 hours, at which timeMS analysis showed that the reaction was completed. The reaction mixturewas concentrated to dryness and then dichloromethane (10 mL) was added.Trifluoroacetic acid was added to this mixture and the mixture wasstirred at room temperature for 4 hours, at which time MS analysisshowed that the reaction was complete. The mixture was then concentratedto half its volume and 1N sodium hydroxide was added to the solutionuntil the pH was adjusted to 14. The organic layer was washed withbrine, then dried over magnesium sulfate and filtered. The filtrate wasconcentrated to give 1.6 g of the title compound as a solid. MS m/z:[M+H⁺] calcd for C₂₀H₂₅N₃O₂ 340.2; found 340.

Preparation 40

[0630]5-[(R)-2-(5-Aminopentylamino)-1-(tert-butyldimethylsilanyloxy)ethyl]-8-benzyloxy-1H-quinolin-2-one

[0631] N-tert-butoxycarbonyl-1,5-diaminopentane (1.04 g, 5.12 mmol) wasadded to a solution of the product of Preparation 13 (1.00 g, 2.05 mmol)in dimethyl sulfoxide (2 mL). The solution was stirred at 75° C. for 12hours, at which time LCMS analysis showed that the reaction wascomplete. The reaction mixture was then concentrated under vacuum todryness. To the residue was added dichloromethane (2 mL) andtrifluoroacetic acid (1 mL) was then added. The solution was stirred atroom temperature for about 3 hours, at which time MS analysis showedthat the reaction was complete. The solution was concentrated to halfits volume and 1N sodium hydroxide was added until the pH was adjustedto 14. The organic layer was collected, washed with brine, dried overmagnesium sulfate and then concentrated to yield 782 mg of the titlecompound as an oil. MS m/z: [M+H⁺] calcd for C₂₉H₄₃N₃O₃Si 510.8; found510.

Preparation 41 Biphenyl-2-ylcarbamic Acid1-[2-(3-{5-[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-(tert-butyldimethylsilanyloxy)ethylamino]pentyl}-ureido)ethyl]piperidin-4-ylEster

[0632] Carbonyl diimidazole (127 mg, 0.78 mmol) was added to a solutionof the product of Preparation 39 (266 mg, 0.78 mmol) in dimethylformamide (4 mL) and the resulting mixture was stirred at roomtemperature for 3 hours. After 3 hours, the product of Preparation 40(399 mg, 0.78 mmol) was added to the reaction mixture and this mixturewas stirred for 12 hours at room temperature, at which time LCMSanalysis determined that the reaction was complete. The reaction mixturewas concentrated in vacuo and the residue was diluted with ethyl acetate(5 mL). The organic layer was washed two times with saturated sodiumbicarbonate (5 mL) and then brine (5 mL). The organic layer was driedover magnesium sulfate, filtered and then concentrated to afford 597 mgof the title compound as a solid which was used without furtherpurification. MS m/z: [M+H⁺] calcd for C₅₀H₆₆N₆O₆Si 875.5; found 875.

Preparation 42 Biphenyl-2-ylcarbamic Acid1-[2-(3-{5-[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethylamino]pentyl}ureido)ethyl]piperidin-4-ylEster

[0633] Triethylamine trihydrofluoride (0.16 mL, 1.02 mmol) was added toa solution of the product of Preparation 41 (597 mg, 0.68 mmol) intetrahydrofuran (3.4 mL) and this mixture was stirred at roomtemperature for about 12 hours, at which time the reaction wasdetermined to be completed by MS analysis. The reaction mixture wasdiluted with ethyl acetate (5 mL) and this mixture was washed with 1Nsodium hydroxide (5 mL), brine, dried over magnesium sulfate andconcentrated to give 417 mg of the title compound as a solid. MS m/z:[M+H⁺] calcd for C₄₄H₅₁N₆O₆ 760.4; found 760.

Example 10 Biphenyl-2-ylcarbamic Acid1-[2-(3-{5-[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]pentyl}ureido)ethyl]piperidin-4-ylEster Ditrifluoroacetate

[0634] A solution of the product of Preparation 42 (417 mg, 0.55 mmol)in ethanol (3 mL) was purged with nitrogen for about 10 minutes.Palladium (10 wt. % (dry basis) on activated carbon) (200 mg) was addedand the solution was flushed again with nitrogen for about 10 minutes.The flask was purged under vacuum and then filled with nitrogen threetimes and then a hydrogen-filled balloon was placed over the flask. Thereaction mixture was stirred under hydrogen for 12 hours, at which timethe reaction was determined to be complete by MS analysis. The reactionmixture was then filtered and the organic filtrate concentrated andpurified by HPLC (10-35% over 60 minute) to give 146 mg of the titlecompound as a powder. MS m/z: [M+H⁺] calcd for C₃₇H₄₆N₆O₆ 671.4; found670. HPLC (10-70) R_(t)=2.6 minutes.

Example 11 Biphenyl-2-ylcarbamic Acid1-[3-(3-{5-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]pentyl}ureido)propyl]piperidin-4-ylEster Ditrifluoroacetate

[0635] Using the method described above in Preparations 39-42 andExample 10, and substituting 3-tert-Butoxycarbonylaminoprop-1-yl bromidefor 2-tert-butoxycarbonylaminoethyl bromide in Preparation 39, the titlecompound was prepared. MS m/z: [M+H⁺] calcd for C₃₈H₄₈N₆O₆ 685.4; found684. HPLC (10 70) R_(t)=2.6 minutes.

Preparation 436-(2-Bromo-(R)-1-tert-butyldimethylsilyloxy)ethyl-2,2-dimethyl-1,3-benzodioxan

[0636] (a) 6-Bromo-2,2-dimethyl-4H-benzo[1,3]dioxine

[0637] To 5-bromo-2-hydroxybenzyl alcohol (93 g, 0.46 mol, availablefrom Sigma-Aldrich) in 2.0 L of 2,2-dimethoxypropane was added 700 mL ofacetone, followed by zinc chloride (170 g). After stirring for 18 hours,1.0 M aqueous sodium hydroxide was added until the aqueous phase wasbasic. Diethyl ether (1.5 L) was added to the slurry and the organicphase was decanted into a separatory funnel. The organic phase waswashed with brine, dried over Na₂SO₄, filtered and concentrated underreduced pressure to give the title compound as an oil.

[0638] (b) 6-Acetyl-2,2-dimethyl-4H-benzo[1,3]dioxine

[0639] To the product of step (a) (110 g, 0.46 mol) in 1.0 L of THF at−78° C. was added 236 mL (0.51 mol) of 2.14 M n-butyllithium in hexanesvia a dropping funnel. After 30 minutes, N-methyl-N-methoxy acetamide(71 g, 0.69 mol, available from TCI) was added. After 2 hours, thereaction mixture was quenched with water, diluted with 2.0 L of 1.0 Maqueous phosphate buffer (pH=7.0) and extracted once with diethyl ether.The diethyl ether phase was washed once with brine, dried over Na₂SO₄,filtered and concentrated under reduced pressure to give a light orangeoil. The oil was dissolved in a minimum volume of ethyl acetate, dilutedwith hexanes, and to give the title compound as a crystalline solid.

[0640] (c) 6-Bromoacetyl-2,2-dimethyl-4H-benzo[1,3]dioxine

[0641] To the product of step (b) (23.4 g, 0.113 mol) in 600 mL of THFat −78° C. was added 135 mL of 1.0 M sodium hexamethyldisilazane in THF(Sigma-Aldrich). After 1 hour, trimethylsilyl chloride (15.8 mL, 0.124mol) was added. After another 30 minutes, bromine (5.82 mL, 0.113 mol)was added. After 10 minutes, the reaction was quenched by diluting thereaction mixture with diethyl ether and pouring it onto 500 mL of 5%aqueous Na₂SO₃ premixed with 500 mL of 5% aqueous NaHCO₃. The phaseswere separated and the organic phase was washed with brine, dried overNa₂SO₄, filtered and concentrated under reduced pressure to give thetitle compound as an oil that solidified upon storage in the freezer.

[0642] (d) (R)-2-Bromo-1-(2,2-dimethyl-4H-benzo[1,3]dioxin-6-yl)ethanol

[0643] To the product of step (c) (10 g, 35.1 mmol) in 100 mL of THF wasadded the solid catalyst of Preparation 13, step (c)(1) (0.97 g, 3.5mmol). The solution was cooled to between −20° C. and −10° C. andBH₃-THF (35 mL, 35 mmol) diluted with 50 mL THF was added dropwise via adropping funnel. After the addition was complete, the reaction mixturewas allowed to warm to ambient temperature. After 30 minutes, thereaction mixture was quenched by slow addition of 50 mL of methanol andthen concentrated to a thick oil. The oil was purified by silica gelchromatography eluted with 1:2 ethyl acetate/hexanes. The fractions werecombined and concentrated to give the title compound as an off-whitesolid.

[0644] (e)[(R)-2-Bromo-1-(2,2-dimethyl-4H-benzo[1,3]dioxin-6-yl)ethoxy]-tert-butyldimethylsilane

[0645] To the product of step (d) (10 g, 34.8 mmol) and imidazole (4.7g, 69.7 mmol) dissolved in 100 mL DMF was added tert-butyldimethylsilylchloride (5.78 g, 38.3 mmol). The reaction mixture was stirred for 18hours. The reaction mixture was then partitioned between 200 mL ofsaturated sodium chloride and 200 mL of diethyl ether. The aqueous layerwas extracted with 200 mL of diethyl ether. The organic layers were thencombined, washed with saturated sodium chloride (3×100 mL), dried overMgSO₄ and concentrated. The product was purified by silica gelchromatography, eluting with hexanes followed by 5% ethyl acetate inhexanes. The desired fractions were combined and concentrated to givethe title compound as an oil.

Preparation 44 Biphenyl-2-ylcarbamic Acid1-{9-[2-(tert-Butyldimethylsilanyloxy)-2-(2,2-dimethyl-4H-benzo[1,3]dioxin-6-yl)ethylamino]nonyl}piperidin-4-ylEster

[0646] The product of Preparation 43 (802 mg, 2.00 mmol) and sodiumiodide (300 mg, 2.00 mmol) were stirred in tetrahydrofuran (0.77 mL) for15 min at ambient temperature. The product of Preparation 11 (675 mg,1.54 mmol) and sodium bicarbonate (388 mg, 4.62 mmol) were added and thereaction mixture was heated at 80° C. for 24 h. The reaction mixture wasthen cooled and water (2 mL) was added. The mixture was then extractedwith dichloromethane (2×2 mL). The combined organic extracts were dried(magnesium sulfate) and the solvent was removed under reduced pressure.The crude residue was purified by flash chromatography (5-10%methanol/dichloromethane) to give the title compound as a solid (798 mg,1.05 mmol, 60% yield). MS m/z: [M+H⁺] calcd for C₄₅H₆₇N₃O₅Si 758.5;found 758.6.

Preparation 45 Biphenyl-2-ylcarbamic Acid1-{9-[2-(2,2-Dimethyl-4H-benzo[1,3]dioxin-6-yl)-2-hydroxyethylamino]nonyl}piperidin-4-ylEster

[0647] Triethylamine trihydrofluoride (342 μL, 2.10 mmol) was added to astirred solution of the Product of Preparation 44 (798 mg, 1.05 mmol) indichloromethane (10.5 mL) at ambient temperature. The reaction mixturewas stirred for 24 h and it was then diluted with dichloromethane (20mL) and washed with saturated aqueous sodium bicarbonate (15 mL). Theorganic layer was dried (magnesium sulfate) and the solvent was removedunder reduced pressure. The crude title compound was isolated as an oil(659 mg, 1.02 mmol), which was used in the next step without furtherpurification. MS m/z: [M+H⁺] calcd for C₃₉H₅₃N₃O₅ 644.4; found 644.8.

Example 12 Biphenyl-2-ylcarbamic Acid1-{9-[(R)-2-Hydroxy-2-(4-hydroxy-3-hydroxymethylphenyl)ethylamino]nonyl}piperidin-4-ylEster Ditrifluoroacetate

[0648] Trifluoroacetic acid (2.80 mL) was added to a stirred solution ofthe product of Preparation 45 (600 mg, 0.93 mmol) in THF/H₂O (14 mL,1:1) and the reaction mixture was stirred for 2 h at ambienttemperature. The reaction mixture was concentrated under reducedpressure and dissolved in 20% MeCN/H₂O then purified by preparative HPLCto yield the title compound (200 mg, 2TFA salt). HPLC (10-70)R_(t)=2.76; MS m/z: [M+H⁺] calcd for C₃₆H₄₉N₃O₅ 604.4; found 604.8.

Preparation 461-[1-(9-Benzylaminononyl)piperidin-4-yl]-3-biphenyl-2-ylurea

[0649] N-Benzylamine (0.903 ml, 8.30 mmol) was added to a solution ofthe product of Preparation 4 (2.40 g, 5.52 mmol) in methanol (25 mL) andthe resulting mixture was stirred at ambient temperature. After 10 min,sodium triacetoxyborohydride (1.75 g, 8.30 mmol) was added to thereaction mixture. The progress of the reaction was followed by HPLCanalysis. After 2 h at ambient temperature, the reaction was quenchedwith water (5 mL) and then concentrated to half its volume under vacuum.The reaction mixture was diluted with dichloromethane (15 mL) and washedwith 1N sodium hydroxide (2×10 mL) and then brine (5 mL). The organiclayer was dried over magnesium sulfate and concentrated to yield thetitle compound.

Preparation 47 2-Benzyloxy-5-(2-bromoacetyl)benzoic Acid Methyl Ester

[0650] (a) 2-Benzyloxy-5-acetylbenzoic Acid Methyl Ester

[0651] Methyl 5-acetylsalicylate (100 g, 0.515 mol) was dissolved inacetonitrile (1 L) in a 2 L flask under reflux conditions and a nitrogenatmosphere. Potassium carbonate (213.5 g, 1.545 mol) was addedportion-wise over 15 min. Benzyl bromide (67.4 mL, 0.566 mol) was addedusing a dropping funnel over 15 min. The reaction was heated to 85° C.for 9 h, and then filtered and rinsed with acetonitrile (100 mL). Thesolution was concentrated to about 300 mL volume under reduced pressureand partitioned between water (1 L) and ethyl acetate (1 L). The organiclayer was washed with saturated sodium chloride (250 mL), dried usingmagnesium sulfate (75 g), and then filtered and rinsed with ethylacetate (100 mL). The organic layer was concentrated to give2-benzyloxy-5-acetylbenzoic acid methyl ester as a solid (100% yield).

[0652] (b) 2-Benzyloxy-5-(2-bromoacetyl)benzoic Acid Methyl Ester

[0653] The product of step (a) (10.0 g, 35.2 mmol) was dissolved inchloroform (250 mL) in a 500 mL flask under a nitrogen atmosphere.Bromine (1.63 mL, 31.7 mmol) dissolved in chloroform (50 mL) was addedusing a dropping funnel over 30 min. The reaction mixture was stirredfor 2.5 h and then concentrated to give a solid. The solid was dissolvedin toluene (150 mL) with some gentle heat, followed by the addition ofethyl ether (150 mL) to yield the title compound as a crystalline solid(55% yield).

Preparation 485-[2-(Benzyl-{9-[4-(3-biphenyl-2-ylureido)piperidin-1-yl]nonyl}amino)acetyl]-2-benzyloxybenzoicAcid Methyl Ester

[0654] The product of Preparation 47 (371 mg, 1.00 mmol) was added to asolution of the product of Preparation 46 (448 mg, 0.85 mmol) indimethyl sulfoxide (4.5 mL) followed by the addition of potassiumcarbonate (234 mg, 1.7 mmol). The reaction mixture was stirred at 40° C.for 6 h, at which time the product of Preparation 46 was no longerobserved by HPLC analysis. The reaction mixture was cooled to ambienttemperature and filtered, and then diluted with ethanol (4 mL). Sodiumborohydride (63 mg, 1.7 mmol) was added to the reaction mixture and thereaction was stirred at ambient temperature for 24 h. The reactionmixture was quenched with 0.5 M ammonium chloride (5 mL) and extractedinto ethyl acetate (2×10 mL). The combined organic layers were washedwith saturated sodium bicarbonate (10 mL) and then with brine (5 mL).The organic layer was dried over magnesium sulfate and the solvent wasremoved under reduced pressure. The crude residue was purified bychromatography on silica gel (3% methanol in chloroform) to give thetitle compound.

Preparation 491-[1-(9-{Benzyl-[2-(4-benzyloxy-3-hydroxymethylphenyl)-2-hydroxyethyl]amino}nonyl)piperidin-4-yl]-3-biphenyl-2-ylurea

[0655] A solution of the product of Preparation 48 (163 mg, 0.20 mmol)in tetrahydrofuran (1.00 mL) was cooled to 0° C. Lithium aluminiumhydride (1.0 M in THF; 0.50 mL, 0.50 mmol) was added dropwise to themixture. After 1 h, the reaction mixture was quenched with water (1 mL)and diluted with ethyl acetate (2 mL). The organic layer was washed withbrine, dried over magnesium sulfate, and the organic extracts werecombined and concentrated to give the title compound.

Example 131-Biphenyl-2-yl-3-(1-{9-[2-hydroxy-2-(4-hydroxy-3-hydroxymethylphenyl)ethylamino]nonyl}piperidin-4-yl)ureaDihydrochloride

[0656] A solution of the product of Preparation 49 (130 mg, 0.16 mmol)in isopropanol (0.80 ml) was flushed with nitrogen for ten minutes andthen palladium (10 wt. % (dry basis) on activated carbon (60 mg) wasadded. The reaction flask was purged with nitrogen and then a balloonfilled with hydrogen was attached to the flask and the reaction mixturewas stirred under an atmosphere of hydrogen. After 72 h, the reactionmixture was filtered and concentrated and the residue was purified bypreparative HPLC. The resulting ditrifluoroacetic acid salt of the titlecompound was dissolved in 1N hydrochloric acid (5 mL) and lyophilized toyield the title compound as its dihydrochloride salt.

Preparation 505-[(R)-2-[(3-Aminomethylcyclohexylmethyl)amino]-1-(tert-butyldimethylsilanyloxy)ethyl]-8-benzyloxy-1H-quinolin-2-one

[0657] A stirred solution of the product of Preparation 13 (1.46 g, 3mmol) and 1,3-cyclohexanebis(methylamine) (426 mg, 3 mmol) in DMSO (3mL) was heated at 100° C. for 6 h. The reaction mixture was allowed tocool and it was then diluted with dichloromethane (20 mL) and washedwith saturated aqueous sodium bicarbonate solution (10 mL). The organiclayer was dried (MgSO₄) and the solvent was removed under reducedpressure. The crude residue was purified by flash chromatography (10%MeOH/DCM and 0.5% NH₄OH) to give the title compound as a solid (775 mg,50% yield). MS m/z: [M+H⁺] calcd for C₃₂H₄₇N₃O₃Si 550.3; found 550.6.

Preparation 51 Biphenyl-2-ylcarbamic Acid1-{2-[(3-{[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydro-quinolin-5-yl)-2-(tert-butyldimethylsilanyloxy)ethylamino]methyl}-cyclohexylmethyl)carbamoyl]ethyl}piperidin-4-ylEster

[0658] To a stirred solution of the product of Preparation 50 (552 mg,1.01 mmol), the product of Preparation 20 (309 mg, 0.84 mmol) and HATU(384 mg, 1.01 mmol) in DMF (1.68 mL) was added DIPEA (190 μL, 1.09mmol). The reaction mixture was stirred at 50° C. overnight and then thesolvent was removed under reduced pressure. The resulting residue wasdissolved in dichloromethane (20 mL) and washed with saturated aqueoussodium bicarbonate solution (10 mL). The organic phase was dried(magnesium sulfate) and the solvent was removed under reduced pressure.The crude product was purified by flash chromatography (5-10% MeOH/DCM)to give the title compound as a solid (267 mg, 36% yield). LCMS (10-70)R_(t)=5.04. MS m/z: [M+H⁺] calcd for C₅₃H₆₉N₅O₆Si 900.5; found 900.6.

Preparation 52 Biphenyl-2-ylcarbamic Acid1-{2-[(3-{[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethylamino]methyl}cyclohexylmethyl)carbamoyl]ethyl}piperidin-4-ylEster

[0659] To a stirred solution of the product of Preparation 51 (267 mg,0.30 mmol) in dichloromethane (3 mL) was added triethylaminetrihydrofluoride (98 μL, 0.6 mmol). The reaction mixture was stirred for10 h and then it was diluted with dichloromethane (10 mL) and washedwith saturated aqueous sodium bicarbonate solution (5 mL). The organicphase was dried (magensium sulfate) and the solvent was removed underreduced pressure to give the title compound as a solid (236 mg, 100%yield). MS m/z: [M+H⁺] calcd for C₄₇H₅₅N₅O₆ 786.4; found 786.5.

Example 14 Biphenyl-2-ylcarbamic Acid1-{2-[(3-{[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-cyclohexylmethyl)carbamoyl]ethyl}-piperidin-4-ylEster

[0660] Palladium (10 wt. % (dry basis) on activated carbon) (120 mg) wasadded to a stirred solution of the product of Preparation 52 (236 mg,0.30 mmol) in ethanol (3 mL). The reaction mixture was placed under ahydrogen atmosphere and stirred overnight. The reaction mixture was thenfiltered and the solvent was removed under reduced pressure. The cruderesidue was purified by preparative HPLC to give the title compound (27mg, 2 TFA salt). HPLC (10-70) R_(t)=2.76. MS m/z: [M+H⁺] calcd forC₄₀H₄₉N₅O₆ 696.4; found 696.6.

Preparation 53 Biphenyl-2-ylcarbamic Acid1-{2-[((1R,3S)-3-Aminocyclopentanecarbonyl)amino]-ethyl}piperidin-4-ylEster

[0661] To a stirred solution of the product of Preparation 39 (318 mg,0.94 mmol), (1R,3S)-3-tert-butoxycarbonylaminocyclopentanecarboxylicacid (258 mg, 1.1 mmol) and HATU (428 mg, 1.1 mmol) in DMF (5 mL) wasadded DIPEA (245 μL, 1.09 mmol). The reaction mixture was stirred atroom temperature overnight and then the solvent was removed underreduced pressure. The resulting residue was dissolved in dichloromethane(20 mL) and washed with saturated aqueous sodium bicarbonate solution(10 mL). The organic layer was dried (magnesium sulfate) and the solventwas removed under reduced pressure. The crude product was purified byflash chromatography (5-10% MeOH/DCM) and then dissolved in atrifluoroacetic acid/DCM mixture (1 mL/5 mL) and stirred at roomtemperature for 1 h. The solvent was removed under reduced pressure. Theresidue was dissolved in dichloromethane (20 mL) and washed with 1Msodium hydroxide (10 mL), dried (magnesium sulfate) and the solventreduced to yield the title compound (167 mg, 39% yield).

Preparation 54 Biphenyl-2-ylcarbamic Acid1-[2-({(1R,3S)-3-[(R)-2-(8-benzyloxy-2-oxo-1,2-dihydroquinoin-5-yl)-2-(tert-butyldimethylsilanyloxy)ethylamino]-cyclopentanecarbonyl}amino)ethyl]piperidin-4-ylEster

[0662] A stirred solution of the product of Preparation 53 (167 mg, 0.38mmol) and the product of Preparation 13 (92 mg, 0.19 mmol) in DMSO (0.38mL) was heated at 90° C. for 5 h. The solution was cooled and dilutedwith ethyl acetate (10 mL) and then washed with saturated aqueous sodiumbicarbonate (5 mL). The organic phase was dried (magnesium sulfate) andthe solvent was removed under reduced pressure. The crude product waspurified by flash chromatography (5-10% MeOH/DCM) to yield the titlecompound (343 mg, 100% yield). LCMS (10-70) R_(t)=4.97. MS m/z: [M+H⁺]calcd for C₅₀H₆₃N₅O₆Si 858.5; found 858.8.

Preparation 55 Biphenyl-2-ylcarbamic Acid1-[2-({(1R,3S)-3-[(R)-2-(8-benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethylamino]cyclopentanecarbonyl}amino)ethyl]-piperidin-4-ylEster

[0663] To a stirred solution of the product of Preparation 54 (343 mg,0.4 mmol) in THF (2 mL) was added triethylamine trihydrofluoride (130μL, 0.8 mmol). The reaction mixture was stirred for 10 h and was thendiluted with EtOAc (10 mL). The reaction mixture was washed withsaturated aqueous sodium bicarbonate solution (5 mL) and then theorganic phase was dried (magnesium sulfate) and the solvent was removedunder reduced pressure to give the title compound as a solid (298 mg,100% yield). HPLC (10-70) R_(t)=2.8. MS m/z: [M+H⁺] calcd for C₄₄H₄₉N₅O₆744.4; found 744.4.

Example 15 Biphenyl-2-ylcarbamic Acid1-[2-({(1R,3S)-3-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2,3,4-tetrahydroquinolin-5-yl)ethylamino]-cyclopentanecarbonyl}amino)ethyl]-piperidin-4-ylEster Ditrifluoroacetate

[0664] To a stirred solution of the product of Preparation 55 (236 mg,0.40 mmol) in ethanol (3 mL) was added palladium (10 wt. % (dry basis)on activated carbon (120 mg). The reaction mixture was placed under ahydrogen atmosphere and stirred overnight. The reaction mixture wasfiltered and the solvent was removed under reduced pressure. The cruderesidue was purified by preparative HPLC to give the title compound (3mg, 2 TFA salt). HPLC (5-75) R_(t)=2.18. MS m/z: [M+H⁺] calcd forC₃₇H₄₅N₅O₆ 656.3; found 656.2.

Preparation 56 4-(tert-Butoxycarbonylaminomethyl)-2-chlorophenylamine

[0665] A stirred solution of 4-aminomethyl-2-chlorophenylamine (940 mg,6 mmol) and di-tert-butyl dicarbonate (1.44 g, 6.6 mmol) indichloromethane (30 mL) was stirred at room temperature for 4 h, atwhich time the reaction was determined to be complete by LCMS. Thereaction mixture was then washed with saturated aqueous sodiumbicarbonate (15 mL) and the organic layer was dried over sodium sulfateand the solvent was removed under reduced pressure. The resulting orangesolid was recrystallized from ethyl acetate to give the titleintermediate as a white solid (˜100% yield).

Preparation 57N-[4-(tert-Butoxycarbonylaminomethyl)-2-chlorophenyl]acrylamide

[0666] To a stirred solution of the product of Preparation 56 (1.54 g,6.0 mmol) in a mixture of diethyl ether (35 mL) and 1 M sodium hydroxide(35 mL) was added dropwise acryloyl chloride (687 μL, 8.45 mmol). After1 h, the organic layer was separated, dried (Na₂SO₄) and the solvent wasremoved under reduced pressure to give the title intermediate as a whitesolid (1.8 g, 96% yield).

Preparation 58 Biphenyl-2-ylcarbamic Acid1-[2-(4-(tert-Butoxycarbonylaminomethyl)-2-chlorophenylcarbamoyl)ethyl]piperidin-4-ylEster

[0667] A solution of the product of Preparation 8 (1.04 g, 3.5 mmol) andthe product of Preparation 57 (1.19 g, 3.85 mmol) in a mixture ofdichloromethane and methanol (12 mL, 1:1) was heated at 60° C. for 12 h.The reaction mixture was allowed to cool and the solvent was removedunder reduced pressure. The crude material was purified by columnchromatography (5-10% MeOH/DCM) to give the title intermediate as awhite solid (2.00 g, 94% yield).

Preparation 59 Biphenyl-2-ylcarbamic Acid1-[2-(4-Aminomethyl-2-chlorophenylcarbamoyl)ethyl]-piperidin-4-yl Ester

[0668] A solution of the product of Preparation 58 (2.00 g, 3.3 mmol)was stirred in dichloromethane (24 mL) and TFA (8 mL) for 1 h and thenthe solvent was removed under reduced pressure. The crude reactionmixture was dissolved in dichloromethane (30 mL) and washed with 1 Msodium hydroxide (2×30 mL). The organic layer was dried (Na₂SO₄) and thesolvent was removed under reduced pressure to give the titleintermediate as an oily white solid (1.46 g, 88% yield).

Preparation 60 Biphenyl-2-ylcarbamic Acid1-[2-(4-{[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-(tert-butyldimethylsilanyloxy)ethylamino]methyl}-2-chlorophenylcarbamoyl)-ethyl]piperidin-4-ylEster

[0669] A stirred solution of the product of Preparation 59 (1.41 g, 2.79mmol) and the product of Preparation 13 (680 mg, 1.39 mmol) in DMSO(1.39 mL) was heated at 90° C. for 8 h and then cooled to roomtemperature. The reaction mixture was diluted with ethylacetate/chloroform (20 mL, 1/1) and the organic layer was washed withsaturated aqueous sodium bicarbonate (10 mL), dried (Na₂SO₄) and thesolvent removed under reduced pressure. The resulting crude residue waspurified by column chromatography (5-10% MeOH/DCM) to give the titleintermediate as a white solid (1.12 g, 88% yield). MS m/z M+H⁺=914.9.

Preparation 61 Biphenyl-2-yl-Carbamic Acid1-[2-(4-{[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethylamino]methyl}-2-chloro-phenylcarbamoyl)ethyl]piperidin-4-ylEster

[0670] To a stirred solution of the product of Preparation 60 (1.12 g,1.23 mmol) in dichloromethane (12 mL) was added Et₃N.3HF (401 μL, 0.6mmol). The reaction mixture was stirred for 10 h and then diluted withdichloromethane (10 mL). This mixture was washed with saturated aqueoussodium bicarbonate solution (5 mL) and the organic layer dried (Na₂SO₄)and the solvent removed under reduced pressure to give the titleintermediate as a white solid (959 mg, 100% yield). MS m/z M+H⁺=800.5.

Example 16 Biphenyl-2-ylcarbamic Acid1-[2-(2-Chloro-4-{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenylcarbamoyl)ethyl]piperidin-4-ylEster Ditrifluoroacetate

[0671] To a stirred solution of the product of Preparation 61 (959 mg,1.2 mmol) in ethanol (12 mL) was added Pd/C (290 mg) and the reactionmixture was placed under a hydrogen atmosphere and stirred overnight.The reaction mixture was then filtered and the solvent removed underreduced pressure. The crude residue was purified by preparative HPLC togive the title compound (67 mg, 2 TFA salt). HPLC (10-70) R_(t)=2.76; MSm/z M+H⁺=710.6.

Preparation 62 2-Chloroethanesulfonic Acid(5-tert-Butoxycarbonylaminopentyl)amide

[0672] To a stirred solution of 5-(tert-butoxycarbonylamino)pentylamine(1.00 g, 4.94 mmol) and triethylamine (689 μL g, 4.94 mmol) indichloromethane (22 mL) at 0° C. was added 2-chloro-1-ethanesulfonylchloride (470 μL, 4.50 mmol). The reaction mixture was stirred for 2 hat room temperature and then washed with saturated aqueous sodiumbicarbonate solution (15 mL). The organic layer was dried (Na₂SO₄) andthe solvent was removed under reduced pressure to give the titlecompound (100% yield), which was used in the next step without furtherpurification.

Preparation 63 Biphenyl-2-ylcarbamic Acid1-[2-(5-tert-Butoxycarbonylaminopentylsulfamoyl)-ethyl]piperidin-4-ylEster

[0673] A solution of the product of Preparation 8 (1.33 g, 3.5 mmol) andthe product of Preparation 62 (1.62 g, 4.94 mmol) in dichloromethane andmethanol (22 mL, 1:1) was heated at 60° C. for 5 h. The reaction mixturewas allowed to cool to room temperature and the solvent was removedunder reduced pressure. The crude residue was dissolved indichloromethane (20 mL) and washed with saturated aqueous sodiumbicarbonate solution (10 mL). The organic layer was then dried (Na₂SO₄)and solvent removed under reduced pressure. The crude residue waspurified by column chromatography (5-10% MeOH/DCM) to give the titleintermediate as a white solid (1.6 g, 55%). MS m/z M+H⁺=589.6.

Preparation 64 Biphenyl-2-ylcarbamic Acid1-[2-(5-Aminopentylsulfamoyl)ethyl]piperidin-4-yl Ester

[0674] A solution of the product of Preparation 63 (1.6 g, 2.72 mmol)was stirred in dichloromethane (21 mL) and TFA (7 mL) for 1 h and thenthe solvent was removed under reduced pressure. The crude reactionmixture was dissolved in dichloromethane (30 mL) and washed with 1 Msodium hydroxide (2×30 mL). The organic layer was dried (Na₂SO₄) and thesolvent was then removed under reduced pressure to give the titleintermediate as an oily white solid (1.19 g, 90% yield).

Preparation 65 Biphenyl-2-ylcarbamic Acid1-(2-{5-[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-(tert-butyldimethylsilanyloxy)ethylamino]pentylsulfamoyl}ethyl)piperidin-4-ylEster

[0675] A stirred solution of the product of Preparation 64 (917 mg, 1.88mmol) and the product of Preparation 13 (460 mg, 0.94 mmol) in DMSO(0.92 mL) was heated at 90° C. for 8 h and then cooled to roomtemperature. The reaction mixture was diluted with ethylacetate/chloroform (20 mL, 1/1) and the organic layer was washed withsaturated aqueous sodium bicarbonate solution (10 mL), dried (Na₂SO₄)and the solvent was removed under reduced pressure. The resulting cruderesidue was purified by column chromatography (3-6% MeOH/DCM) to givethe title intermediate as a white solid (500 mg, 60% yield). MS m/zM+H⁺=896.9.

Preparation 66 Biphenyl-2-ylcarbamic Acid1-(2-{5-[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethylamino]pentylsulfamoyl}ethyl)piperidin-4-ylEster

[0676] To a stirred solution of the product of Preparation 65 (500 mg,0.56 mmol) in dichloromethane (5.6 mL) was added triethylaminetrihydrofluoride (183 μL, 1.12 mmol). The reaction mixture was stirredfor 10 h and dichloromethane (10 mL) was added. The resulting mixturewas washed with saturated aqueous sodium bicarbonate solution (5 mL).The organic layer was dried (Na₂SO₄) and the solvent was removed underreduced pressure to give the title intermediate as a yellow solid (437mg, 100% yield. MS m/z M+H⁺=782.8.

Example 17 Biphenyl-2-ylcarbamic Acid1-(2-{5-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]pentylsulfamoyl}ethyl)piperidin-4-ylEster Ditrifluoroacetate

[0677] To a stirred solution of the product of Preparation 66 (437 mg,0.56 mmol) in ethanol/methanol (5.6 mL, 1/1) was added Pd/C (131 mg) andthe reaction mixture placed under a hydrogen atmosphere and stirredovernight. The reaction mixture was then filtered and the solvent wasremoved under reduced pressure. The crude residue was purified bypreparative HPLC to give the title compound as the ditrifluoroaceticacid salt (71 mg). HPLC (10-70) R_(t)=2.59; MS m/z M+H⁺=692.6

Preparation 67 Biphenyl-2-ylcarbamic Acid1-{2-[(4-Formylbenzenesulfonyl)methylamino]-ethyl}piperidin-4-yl Ester

[0678] To a stirred solution of the product of Preparation 26 (350 mg, 1mmol) and triethylamine (167 μL, 1.2 mmol) in dichloromethane (5 mL) wasadded 4-formylbenzenesulfonyl chloride (225 mg, 1.1 mmol). After 1 h atroom temperature, the reaction was complete by MS and the reactionmixture was then washed with saturated aqueous sodium bicarbonatesolution (5 mL). The organic layer was then dried (Na₂SO₄) and solventremoved under reduced pressure to give the title intermediate (323 mg,62% yield). MS m/z M+H⁺=522.4.

Preparation 68 Biphenyl-2-ylcarbamic Acid1-{2-[(4-{[(R)-2-(tert-Butyldimethylsilanyloxy)-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}benzenesulfonyl)-methylamino]ethyl}piperidin-4-ylEster

[0679] A solution of5-[(R)-2-amino-1-(tert-butyldimethylsilanyloxy)ethyl]-8-hydroxy-1H-quinolin-2-one(293 mg, 0.74 mmol) and the product of Preparation 67 in dichloromethaneand methanol (6.2 mL, 1/1) was stirred at room temperature for 1 h andthen sodium triacetoxyborohydride (394 mg, 1.86 mmol) was added. Thereaction mixture was stirred for 4 h at which time the reaction wasdetermined to be complete by MS. The reaction mixture was then acidifiedwith concentrated hydrochloric acid and the solvent was removed underreduced pressure to provide the title compound, which was used in thenext step without further purification. MS m/z M+H⁺=840.8.

Example 18 Biphenyl-2-ylcarbamic Acid1-{2-[(4-{[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}benzenesulfonyl)methylamino]ethyl}-piperidin-4-ylEster Ditrifluoroacetate

[0680] A stirred solution of the product of Preparation 68 (520 mg, 0.62mmol) in 1M hydrochloric acid (5 mL) and acetonitrile (5 mL) was heatedat 60° C. for 8 h. The reaction mixture was cooled to room temperatureand the solvent was removed under reduced pressure. The crude residuewas purified by preparative HPLC to give the title compound as theditrifluoroacetic acid salt (220 mg). HPLC (10-70) R_(t)=2.77; MS m/zM+H⁺=726.7.

Preparation 69 (3-Aminomethylphenyl)methanol Hydrochloride

[0681] (a) (3-tert-Butoxycarbonylmethylphenyl)methanol

[0682] Borane dimethyl sulfide (2.05 mL, 21.6 mmol) was added to asolution of 3-(tert-butoxycarbonylaminomethyl)benzoic acid (1.81 g, 7.20mmol) in tetrahydrofuran (24 mL). and the resulting mixture was stirredat room temperature for 3 hours. The reaction mixture was then dilutedwith ethyl acetate (20 mL) and the layers were separated. The organiclayer was washed with saturated sodium bicarbonate, saturated sodiumchloride, dried over magnesium sulfate and concentrated to give thetitle compound as a yellow oil (1.71 g).

[0683] (b) (3-Aminomethylphenyl)methanol Hydrochloride

[0684] To the product of step (a) (1.71 g, 7.2 mmol) was added asolution of 4 M hydrochloric acid in dioxane (9 mL, 36 mmol) and theresulting mixture was stirred at room temperature for 1 h. The reactionmixture was then concentrated and the residue was diluted with diethylether (50 mL) and filtered to provide the title compound as a whitesolid (1.09 g).

Preparation 70 Biphenyl-2-ylcarbamic Acid1-{2-[3-(3-Hydroxymethylbenzyl)ureido]ethyl}piperidin-4-yl Ester

[0685] A 0.2 M solution of the product of Preparation 35 (760 mg, 2.24mmol) in N,N-dimethylformamide was added dropwise to a solution of1,1′-carbonyldiimidazole (364 mg, 2.24 mmol) and diisopropylethylamine(0.31 mL, 2.24 mmol) in N,N-dimethylformamide (11 mL) and the resultingmixture was stirred at room temperature for 2 h. Diisopropylethylamine(0.31 mL, 2.24 mmol) and the product of Preparation 69 (578 mg, 3.4mmol) were added and this mixture was stirred at 50° C. for 12 hours.The reaction mixture was then concentrated to dryness and the residuewas diluted with dichloromethane (20 mL) and this solution was washedwith saturated sodium bicarbonate (2×), saturated sodium chloride, driedover magnesium sulfate, and concentrated to provide the title compound(1.12 g). LCMS (2-90) R_(t)=4.01 min.; MS m/z M+H=503.5.

Preparation 71 Biphenyl-2-ylcarbamic Acid1-{2-[3-(3-Formylbenzyl)ureido]ethyl}piperidin-4-yl Ester

[0686] A solution of the product of Preparation 70 (1.12 g, 2.23 mmol)in dichloromethane (11.1 mL) was cooled to ⁰° C. anddiisopropylethylamine (1.17 mL, 6.70 mmol) and dimethyl sulfoxide (0.949mL, 13.4 mmol) were added. After about 10 minutes, pyridine sulfurtrioxide complex (1.06 g, 6.70 mmol) was added and the resulting mixturewas stirred at 0° C. for 2 h. The reaction was then quenched with water(15 mL) and the organic layer was washed with cold water (3×), driedover magnesium sulfate and concentrated to provide the title compound asa yellow crisp (609 mg). LCMS (2-90) R_(t)=4.13 min; MS m/z M+H=501.3.

Preparation 72 Biphenyl-2-ylcarbamic Acid1-{2-[3-(3-{[(R)-2-(tert-butyldimethylsilanyloxy)-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}benzyl)ureido]ethyl}-piperidin-4-ylEster

[0687]5-[(R)-2-Amino-1-(tert-butyldimethylsilanyloxy)ethyl]-8-hydroxy-1H-quinolin-2-one(575 mg, 1.40 mmol) was added to a solution of the product ofPreparation 71 (609 mg, 1.2 mmol) and diisopropylamine (0.25 mL, 1.40mmol) in dichloromethane (6 mL) and the resulting mixture was stirred atroom temperature for 45 min. Sodium triactetoxyborohydride (385 mg, 1.80mmol) was then added and this mixture was stirred at room temperaturefor 12 h. The reaction was then quenched with 10% aqueous hydrochloricacid (5 mL) and the layers were separated. The organic layer was washedwith saturated sodium bicarbonate, saturated sodium chloride, dried overmagnesium sulfate and concentrated to give the title compound (1.1 g).HPLC (10-70) R_(t)=3.55 min; MS m/z M+H=819.7.

Example 19 Biphenyl-2-ylcarbamic Acid1-{2-[3-(3-{[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}benzyl)ureido]ethyl}piperidin-4-ylEster Ditrifluoroacetate

[0688] Triethylamine trihydrofluoride (2.4 mL, 13.6 mmol) was added to asolution of the product of Preparation 72 (1.1 g, 1.36 mmol) indichloromethane (2 mL) and the resulting mixture was stirred at roomtemperature for 15 h. The reaction mixture was then concentrated undervacuum to dryness and the residue was dissolved in a 1:1 mixture ofwater and acetonitrile with 0.1% TFA and this mixture was purified byHPLC (5-35 over 60 min) to provide the title compound as theditrifluroacetate salt (296 mg, 99% purity). MS m/z M+H=705.6.

Preparation 73 Biphenyl-2-ylcarbamic Acid1-[(E)-3-(4-Nitrophenyl)allyl]piperidin-4-yl Ester

[0689] The product of Preparation 8 (2.96 g, 0.01 mol) andp-nitrocinnamaldehyde (1.77 g, 0.01 mol) were stirred in 50 mL ofdichloromethane for 2 h. Sodium triacetoxyborohydride (6.33 g, 0.03 mol)was added and the resulting mixture was stirred for 2 h. The reactionwas then quenched with 10 mL of water and this mixture was diluted withdichloromethane (100 mL). The organic layer was washed with saturatedsodium bicarbonate (2×), brine, dried over Na₂SO₄, filtered andconcentrated to provide the title compound as a yellow foam (3.8 g, 80%yield).

Preparation 74 Biphenyl-2-ylcarbamic Acid1-[3-(4-Aminophenyl)propyl]piperidin-4-yl Ester

[0690] The product of Preparation 73 (2.5 g, 5.4 mmol) was dissolved in100 mL of ethanol and the resulting solution was purged with nitrogenfor 30 min. Palladium on carbon (2.5 g; 50% w/w water; 10% Pd; 1.1 mmolPd) was then added while degassing with nitrogen. This mixture was thenplaced under hydrogen (50 psi) until hydrogen was no longer consumed(˜30 minutes). The mixture was then purged with nitrogen, filteredthrough Celite and concentrated. The residue was dissolved in ethylacetate and this mixture was washed with saturated sodium bicarbonate(2×), brine, dried (Na₂SO₄), filtered and concentrated to provide thetitle compound (2.08 g, 90% yield). MS m/z M+H=430.5.

Preparation 75 Biphenyl-2-ylcarbamic Acid1-{3-[4-(4-{2-[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-(tert-butyldimethylslanyloxy)ethylamino]ethyl}phenylamino)phenyl]propyl}piperidin-4-ylEster

[0691] To a 25 mL round-bottomed flask was added the product ofPreparation 74 (400 mg, 0.8 mmol);8-benzyloxy-5-[(R)-2-[2-(4-bromophenyl)ethylamino]-1-(tert-butyldimethylsilanyloxy)ethyl]-1H-quinolin-2-one(769 mg, 1.2 mmol); tris(dibenzylideneacetone)dipalladium(0) (73 mg,0.08 mmol, 20% Pd); and 2-(dicyclohexylphosphino)biphenyl (84 mg, 0.24mmol). This mixture was purged with nitrogen and then dry, degassedtoluene (8 mL, 0.1 M) was added and the resulting mixture was heated at70° C. for 30 min. Sodium tert-butoxide (382 mg, 4.0 mmol) was thenadded, and the temperature was raised to 95° C. for 4 h, at which timeLCMS showed complete consumption of the product of Preparation 74 and alarge product peak (M+H=956.7). The reaction mixture was then cooled toroom temperature and diluted with ethyl acetate. This mixture was washedwith saturated sodium bicarbonate (2×), brine, dried (Na₂SO₄), filteredand concentrated to provide the title compound (1.5 g), which was usedwithout further purification.

Preparation 76 Biphenyl-2-ylcarbamic Acid1-{3-[4-(4-{2-[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethylamino]ethyl}phenylamino)phenyl]propyl}piperidin-4-ylEster

[0692] The product of Preparation 75 was dissolved in dichloromethane(10 mL) and triethylamine trihydrofluoride (10 eq.) was added. Thereaction mixture was stirred overnight and then diluted withdichloromethane and the organic layer was washed with saturated sodiumbicarbonate (2×), brine, dried (Na₂SO₄), filtered and concentrated toprovide 1.3 g of crude product. This material was purified by silica gelchromatography (DCM, incrementally to 50% methanol) to provide the titlecompound (300 mg, about 75% purity), which was used without furtherpurification.

Example 20 Biphenyl-2-ylcarbamic acid1-{3-[4-(4-{2-[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylamino)phenyl]propyl}piperidin-4-ylester Ditrifluoroacetate

[0693] The product of Preparation 76 (300 mg) was dissolved in 10 mL ofethanol and this mixture was purged with nitrogen for 15 minutes.Palladium on carbon (10% Pd, 50% w/w water, 0.2 eq. Pd) was added whiledegassing. The resulting mixture was then placed under 1 atm. ofhydrogen for 2 h, at which time the reaction was complete by LCMS. Thesolution was then purged with nitrogen for 15 min and then filteredthrough Celite and concentrated. The resulting residue was purified byprep HPLC to afford the title compound as the ditrifluoroacetate salt(59 mg, >95% purity). MS m/z M+H=752.8.

Preparation 77 Biphenyl-2-ylcarbamic Acid1-[2-Fluoro-3-(4-hydroxymethylpiperidin-1-ylmethyl)-benzyl]piperidin-4-ylEster

[0694] The product of Preparation 8 (500 mg, 1.69 mmol),2,6-bis(bromomethyl)-1-fluorobenzene (476 mg, 1.69 mmol,piperidin-4-ylmethanol (195 mg, 1.69 mmol) and potassium carbonate (466mg, 3.37 mmol) were suspended in acetonitrile (5 mL) and stirred at roomtemperature for 18 h. The reaction mixture was then concentrated and theresidue was dissolved in dichloromethane/water. The layers wereseparated and the organic layer was washed with water (2×), brine, dried(MgSO₄) and concentrated. The crude material was purified by silica gelcolumn chromatography eluting with 3% methanol/chloroform to give thetitle compound as a white foam (282 mg). MS m/z M+H=532.3.

Preparation 78 Biphenyl-2-ylcarbamic Acid1-[2-Fluoro-3-(4-formylpiperidin-1-ylmethyl)benzyl]-piperidin-4-yl Ester

[0695] The product of Preparation 77 (282 mg, 0.53 mmol) was dissolvedin dichloromethane and to this mixture was added diisopropylethylamine(280 μL, 1.6 mmol) and dimethyl sulfoxide (115 μL, 1.6 mmol). Thereaction mixture was cooled to −15° C. under nitrogen and pyridinesulfur trioxide complex (255 mg, 1.6 mmol) was added and the resultingmixture was stirred for 40 min. The reaction was then quenched withwater and the layers were separated. The organic layer was washed withaqueous NaH₂PO₄ (1M×3), brine, dried (MgSO₄) and concentrated to providethe title compound as a foam (253 mg). MS m/z M+H=530.4.

Example 21 Biphenyl-2-ylcarbamic Acid1-[2-Fluoro-3-(4-{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}piperidin-1-ylmethyl)benzyl]piperidin-4-ylEster Ditrifluoroacetate

[0696] The product of Preparation 78 (253 mg, 0.48 mmol) was dissolvedin a 1:1 mixture of dichloromethane and methanol (6 mL) and to thismixture was added5-[(R)-2-amino-1-(tert-butyldimethylsilanyloxy)ethyl]-8-hydroxy-1H-quinolin-2-oneacetate (228 mg, 0.58 mmol) and sodium triacetoxyborohydride (317 mg,1.5 mmol). The reaction mixture was stirred under nitrogen at roomtemperature for 18 h and then concentrated. The residue was dissolved ina 2:3 mixture of acetonitrile and aqueous 6 N hydrochloric acid, andthis mixture was heated at 55° C. for 4 hours. The reaction mixture wasthen concentrated and the residue was dissolved inwater/acetonitrile/trifluoroacetic acid (1:1:0.005) and purified byreverse phase column chromatography to afford the title compound as awhite solid (175 mg). MS m/z M+H=734.5.

Preparation 79 2-[4-(3-Bromopropoxy)phenyl]ethanol

[0697] To a solution of 4-hydroxyphenethyl alcohol (4.37 g, 31.0 mmol)and potassium carbonate (6.55 g, 47.0 mmol) in acetonitrile (62.0 mL)was added 1,3 dibromopropane (31.0 mL, 316 mmol). The reaction mixturewas heated to 70° C. for 12 hours and then cooled to room temperature,filtered and concentrated under vacuum. The resulting oil was purifiedby silica gel chromatography using a mixture of 4:1 hexanes and ethylacetate to give the title compound (6.21 g) as a white solid.

Preparation 80 Biphenyl-2-ylcarbamic Acid1-{3-[4-(2-Hydroxyethyl)phenoxy]propyl}piperidin-4-yl Ester

[0698] To a solution of the product of Preparation 79 (1.11 g, 4.30mmol) and diisopropylethylamine (0.90 mL, 5.10 mmol) in acetonitrile(21.5 mL) was added the product of Preparation 8 (1.27 g, 4.30 mmol) andthe resulting mixture was stirred at 60° C. for 12 h. The reactionmixture was then diluted with dichloromethane (20 mL) and washed withsaturated sodium bicarbonate (25 mL), saturated sodium chloride (25 mL),dried over magnesium sulfate and concentrated to provide the titlecompound (1.98 g, 85% purity). MS m/z M+H=475.5.

Preparation 81 Biphenyl-2-ylcarbamic Acid1-{3-[4-(2-Oxoethyl)phenoxy]propyl}piperidin-4-yl Ester

[0699] A solution of the product of Preparation 80 (723 mg, 1.53 mmol)and dichloromethane (75 mL) was cooled to about 5° C. anddiisopropylethylamine (798 mL, 4.58 mmol) and dimethyl sulfoxide (649mL, 9.15 mmol) were added. Pyridine sulfur trioxide (728 mg, 4.58 mmol)was then added and the resulting mixture was stirred at 5° C. for 45min. The reaction mixture was then diluted with dichloromethane (20 mL)and washed with saturated sodium bicarbonate (25 mL), saturated sodiumchloride (25 mL), dried over magnesium sulfate and concentrated toprovide the title compound (604 mg). MS m/z M+H=473.4.

Preparation 82 Biphenyl-2-ylcarbamic Acid1-[3-(4-{2-[(R)-2-(tert-butyldimethylsilanyloxy)-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}-phenoxy)propyl]piperidin-4-ylEster

[0700] The product of Preparation 81 (604 mg, 1.28 mmol) was dissolvedin methanol (6.4 mL) and5-[(R)-2-amino-1-(tert-butyldimethylsilanyloxy)ethyl]-8-hydroxy-1H-quinolin-2-one(605 mg, 1.53 mmol) and diisopropylethylamine (0.27 mL, 1.53 mmol) wereadded. Sodium triacetoxyborohydride (405 mg, 1.91 mmol) was then addedand the reaction mixture was stirred at room temperature for 3 h. Thereaction mixture was then concentrated to dryness and the residue wasdiluted with ethyl acetate (20 mL) and this solution was washed withsaturated sodium bicarbonate (25 mL), saturated sodium chloride (25 mL),dried over magnesium sulfate and concentrated to give the title compound(704 mg). MS m/z M+H=791.8.

Example 22 Biphenyl-2-ylcarbamic Acid1-[3-(4-{2-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenoxy)propyl]piperidin-4-ylEster Ditrifluoroacetate

[0701] Triethylamine trihydrofluoride (1.5 mL, 8.87 mmol) was added to asolution of the product of Preparation 82 (702 mg, 0.89 mmol) indichloromethane (4.5 mL) and the resulting mixture was stirred at roomtemperature for 24 h. The mixture was then concentrated under vacuum andpurified by HPLC (2-35 over 90 min) to give the title compound (92 mg)as a white powder. MS m/z M+H=677.4.

Preparation 83 Methyl 4-Iodophenylacetate

[0702] To a stirred solution of 4-iodophenylacetic acid (5.0 g, 19.1mmol) in MeOH (200 mL) was added 4N hydrochloric acid in dioxane (10mL). The reaction mixture was stirred for 24 h at room temperature andthen the solvent was removed under reduced pressure to give the titlecompound (5.17 g, 98% yield), which was used without furtherpurification.

Preparation 84 Methyl [4-(4-Hydroxybut-1-ynyl)phenyl]acetate

[0703] To a stirred solution of the product of Preparation 83 (4.5 g,16.3 mmol) in diethylamine (100 mL) was added but-3-yn-1-ol (1.9 mL,32.6 mmol), Pd(PPh₃)₂Cl₂ (500 mg, 1.63 mmol) and CuI (154 mg, 0.815mmol) and resulting mixture was stirred for 17 h at room temperature.The solvent was then removed under reduced pressure and the residue wasdissolved in diethyl ether (200 mL) and this solution was filtered toremove salts. The solvent was then removed under reduced pressure andthe crude product was purified by silica gel chromatography (60%EtOAc/Hexane) to afford the title intermediate (3.03 g, 91% yield).

Preparation 85 Methyl [4-(4-Hydroxybutyl)phenyl]acetate

[0704] A stirred solution of the product of Preparation 84 (2.8 g, 12.8mmol) in methanol (50 mL) was flushed with nitrogen and then 10%palladium on carbon (400 mg, 20% wt/wt) was added. The reaction flaskwas then alternately placed under vacuum and flushed with hydrogen forcycles and then stirred under hydrogen for 14 h. The reaction mixturewas flushed with nitrogen and then filtered and the solvent removedunder reduced pressure to give the title compound (2.75 g, 97% yield),which was used without further purification.

Preparation 86 Methyl(4-{4-[4-(Biphenyl-2-ylcarbamoyloxy)piperidin-1-yl]butyl}phenyl)acetate

[0705] (a) Methyl {4-[4-(Toluene-4-sulfonyloxy)butyl]phenyl}acetate

[0706] To a stirred solution of the product of Preparation 85 (2.6 g,12.5 mmol) in THF (100 mL) was added DABCO (2.6 g, 25.0 mmol) and thenp-toluenesulfonyl chloride (2.44 g, 13.75 mmol). The reaction mixturewas stirred at room temperature for 23 h and then solvent was removedunder reduced pressure and the residue was dissolved in dichloromethane(200 mL). The organic layer was then washed with water (2×100 mL), 1Nhydrochloric acid (100 mL), aqueous saturated sodium chloride solution(100 mL), dried (MgSO₄), filtered and the solvent removed under reducedpressure to give the title compound, which was used without furtherpurification.

[0707] (b) Methyl(4-{4-[4-(Biphenyl-2-ylcarbamoyloxy)piperidin-1-yl]butyl}phenyl)acetate

[0708] To the crude product from step (a) was added DMF (50 mL),diisopropylethylamine (3.0 mL, 17.3 mmol) and the product of Preparation8 (2.4 g, 8.1 mmol). The reaction mixture was stirred at roomtemperature for 18 h and then the solvent was removed under reducedpressure to give the title compound (3.5 g, 86.3% yield). MS m/z 501.6(MH⁺), R_(f) 4.89 min (10-70% ACN: H₂O, reverse phase HPLC).

Preparation 87 Biphenyl-2-ylcarbamic Acid1-{4-[4-(2-Hydroxyethyl)phenyl]butyl}piperidin-4-yl Ester

[0709] To a stirred solution of the product of Preparation 86 (2.0 g,4.0 mmol) in THF (100 mL) was added dropwise DIBAL (24 mL, 24 mmol, 1.0M in THF). After the addition was complete, the reaction mixture wasstirred for 3 h and then quenched by slow addition of methanol (untilgas evolution ceased). The mixture was then stirred for 30 min. and thenethyl acetate (200 mL) and aqueous 1N sodium hydroxide (200 mL) wereadded. The organic layer was separated and washed with aqueous saturatedsodium chloride solution (100 mL), dried (MgSO₄), filtered and thesolvent removed under reduced pressure to give the title compound (1.3g, 69% yield), which was used without further purification. MS m/z 473.4(MH⁺), R_(f) 4.53 min (10-70% ACN: H₂O, reverse phase HPLC).

Example 23 Biphenyl-2-ylcarbamic Acid1-[2-(4-{2-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenyl)ethyl]piperidin-4-ylEster

[0710] To a stirred solution of the product of Preparation 87 (500 mg,1.06 mmol) in dichloromethane (25 mL) was added dimethyl sulfoxide (0.60mL, 10.6 mmol) and diisopropylethylamine (0.921 mL, 5.3 mmol). Thereaction mixture was then cooled to −10° C. and pyridine sulfur trioxide(842 mg, 5.3 mmol) was added. The reaction mixture was stirred for 1 hand then quenched by adding water (100 mL). This mixture was stirred for10 min and then the organic layer was removed and washed with aqueoussaturated sodium chloride solution (100 mL), dried (MgSO₄) and thenfiltered.

[0711] To the filtrate was added methanol (25 mL),5-[(R)-2-amino-1-(tert-butyldimethylsilanyloxy)ethyl]-8-hydroxy-1H-quinolin-2-oneacetate (419 mg, 1.06 mmol) and sodium triacetoxyborohydride (468 mg,2.12 mmol). This mixture was stirred for 16 h and then condensed and, tothe resulting mixture, was added a 1:1 mixture of acetonitrile andaqueous 4N hydrochloric acid (20 mL). This mixture was heated at 50° C.for 17 h and then the solvent was removed under reduced pressure. To theresidue was added a 1:1 mixture of acetic acid and water (8.0 mL) andthe mixture was chromatographed on reverse-phase silica gel (gradientelution, 10-50% ACN/H₂O) to afford the title compound (67 mg, 7% yieldover 3 steps). MS m/z (MH⁺) 675.5; R_(f) 3.07 (10-70% ACN: H₂O, reversephase HPLC).

Preparation 88 Ethyl 3-[5-(2-Ethoxycarbonylvinyl)thiophen-2-yl]acrylate

[0712] To a stirred solution of sodium hydride (2.1 g, 53 mmol, 60% inmineral oil) in THF (200 mL) was slowly added triethylphosphonoacetate(10 mL, 50 mmol) Hydrogen gas evolution was observed and the reactionwas stirred until gas evolution ceased (about 30 min). To this reactionmixture was added 2,5-thiophenedicarboxaldehyde (3 g, 21 mmol) and thereaction mixture was stirred for 1 h. The solvent was removed underreduced pressure and the residue was dissolved in dichloromethane (200mL). The organic layer was washed with water (100 mL), aqueous 1Nhydrochloric acid (100 mL), aqueous saturated sodium chloride solution(100 mL), dried (MgSO₄), filtered and the solvent removed under reducedpressure to give the title compound (5.8 g, 98% yield), which was usedwithout further purification.

Preparation 89 Ethyl3-[5-(2-Ethoxycarbonylethyl)thiophen-2-yl]propionate

[0713] A stirred solution of the product of Preparation 88 (5.8 g, 21mmol) in methanol (200 mL) was flushed with nitrogen and 10% palladiumon carbon (576 mg, 10% wt/wt) was added. The reaction flask wasalternately placed under vacuum and flushed with hydrogen for 3 cyclesand then the reaction mixture was stirred under hydrogen for 1 h. Themixture was then was flushed with nitrogen, filtered and the solventremoved under reduced pressure to give the title compound (5.8 g, 99%yield), which was used without further purification.

Preparation 90 3-[5-(3-Hydroxypropyl)thiophen-2-yl]propan-1-ol

[0714] To a stirred solution of DIBAL (88 mL, 88 mmol, 1.0M incyclohexane) in THF (300 mL) at −78° C. was added dropwise the productof Preparation 89 (5.0 g, 17.6 mmol). After the addition was complete,the reaction mixture was warmed to room temperature over 30 min and thenquenched by slow addition of aqueous 1N hydrochloric acid (200 mL).Dichloromethane (400 mL) was added and the layers were separated. Theaqueous layer was washed with dichloromethane (4×100 mL) and thecombined organic layers were washed with aqueous saturated sodiumchloride solution (100 mL), dried (MgSO₄), filtered and the solventremoved under reduced pressure to give the title compound (3.0 g, 85%yield), which was used without further purification.

Preparation 91 Biphenyl-2-ylcarbamic Acid1-{3-[5-(3-Hydroxypropyl)thiophen-2-yl]propyl}piperidin-4-yl Ester

[0715] (a) Toluene-4-sulfonic Acid3-[5-(3-Hydroxypropyl)thiophen-2-yl]propyl Ester

[0716] To a stirred solution of the product of Preparation 90 (423 mg,2.1 mmol) in THF (20 mL) was added DABCO (420 mg, 4.2 mmol) and thenp-toluenesulfonyl chloride (442 mg, 2.3 mmol). The reaction mixture wasstirred at room temperature for 2 h and then the solvent was removedunder reduced pressure and the residue was dissolved in dichloromethane(200 mL). The organic layer was washed with water (2×100 mL), aqueoussaturated sodium chloride solution (100 mL), dried MgSO₄), filtered andthe solvent removed under reduced pressure to give the title compound,which was used without further purification.

[0717] (b) Biphenyl-2-ylcarbamic Acid1-{3-[5-(3-Hydroxypropyl)thiophen-2-yl]propyl}piperidin-4-yl Ester

[0718] To the product from step (a) was added acetonitrile (20 mL),diisopropylethylamine (0.5 mL, 2.8 mmol) and the product of Preparation8 (626 mg, 2.11 mmol). The reaction mixture was heated to 50° C. for 20h and then cooled to room temperature and the solvent was removed underreduced pressure. The residue was purified by silica gel chromatography(5% MeOH/DCM with 0.6% NH₃ (aq)) to afford the title compound (450 mg,44% yield). MS m/z (MH⁺) 479.6; R_(f) 4.15 min (10-70% ACN: H₂O, reversephase HPLC).

Preparation 92 Biphenyl-2-ylcarbamic Acid1-[3-(5-{3-[(R)-2-(tert-Butyldimethylsilanyloxy)-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]propyl}thiophen-2-yl)propyl]-piperidin-4-ylEster

[0719] To a stirred solution of the product of Preparation 91 (450 mg,0.94 mmol) in dichloromethane (20 mL) was added dimethyl sulfoxide (0.21mL, 3.7 mmol) and diisopropylethylamine (0.65 mL, 3.7 mmol). Thismixture was cooled to −10° C. and pyridine sulfur trioxide (444 mg, 2.8mmol) was added. The reaction mixture was stirred for 3 h and then wasquenched by adding water (100 mL). This mixture was stirred for 10 minand then the organic layer was removed and was washed with aqueoussaturated sodium chloride solution (100 mL), dried (MgSO₄) and filtered.

[0720] To the filtrate was added methanol (20 mL),5-[(R)-2-amino-1-(tert-butyldimethylsilanyloxy)ethyl]-8-hydroxy-1H-quinolin-2-oneacetate (368 mg, 0.93 mmol) and then sodium triacetoxyborohydride (412mg, 1.86 mmol). This mixture was stirred for 19 h and then the mixturewas condensed to give the title compound, which was used without furtherpurification. MS m/z (MH⁺) 795.8; R_(f) 4.93 min (10-70% ACN: H₂O,reverse phase HPLC).

Example 24 Biphenyl-2-ylcarbamic Acid1-[3-(5-{3-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]propyl}thiophen-2-yl)propyl]piperidin-4-ylEster

[0721] To the crude product from Preparation 92 was added a 1:1 mixtureof acetonitrile and aqueous 4 N hydrochloric acid (25 mL). This mixturewas heated at 50° C. for 17 h and then the solvent was then removedunder reduced pressure. To the residue was added a 1:1 mixture of aceticacid and water (8.0 mL) and this mixture was chromatographed onreverse-phase silica gel (gradient elution, 10-50% ACN/H₂O) to affordthe title compound (135 mg, 16% yield for 3 steps). MS m/z (MH⁺) 681.5;R_(f) 3.03 (10-70% ACN: H₂O, reverse phase HPLC).

Preparation 93 Methyl 4-Amino-5-chloro-2-methoxybenzoate

[0722] To a solution of 4-amino-5-chloro-3-methoxybenzoic acid (1.008 g,5.0 mmol) in a mixture of toluene (9 mL) and methanol (1 mL) at 0° C.was added (trimethylsilyl)diazomethane (2.0 M in hexane, 3.0 mL, 6.0mmol) dropwise. The reaction mixture was then warmed to room temperatureand stirred for 16 h. Excess (trimethylsilyl)diazomethane was quenchedby adding acetic acid until the bright yellow color of the reactionmixture disappeared. The mixture was then concentrated in vacuo to givethe title compound as an off-white solid, which was used without furtherpurification.

Preparation 94 Methyl 4-Acryloylamino-5-chloro-2-methoxybenzoate

[0723] To crude product of Preparation 93 was added dichloromethane (10mL, 0.5 M) and triethylamine (2.1 mL, 15 mmol). This mixture was cooledto 0° C. and acryloyl chloride (812 μL, 10 mmol) was added dropwise withstirring. After 2 h, the reaction was quenched by adding methanol (about2 mL) at 0° C. and the resulting mixture was stirred at room temperaturefor 15 min and then concentrated in vacuo. Dichloromethane (30 mL) andwater (30 mL) were added to the residue and this mixture was mixedthoroughly. The layers were separated and the aqueous layer wasextracted with dichloromethane (20 mL). The organic layers werecombined, dried (Na₂SO₄), filtered and the solvent was removed in vacuoto give the title compound as a brown foamy solid, which was usedwithout further purification.

Preparation 95 Methyl4-{3-[4-(Biphenyl-2-ylcarbamoyloxy)piperidin-1-yl]propionylamino}-5-chloro-2-methoxybenzoate

[0724] To the crude product from Preparation 94 was added the product ofPreparation 8 (1.33 g, 4.5 mmol) and a mixture of THF (22.5 mL) andmethanol (2.5 mL). This mixture was heated at 50° C. with stirring for16 h and then the solvent was removed in vacuo. The residue waschromatographed (silica gel; EtOAc) to give the title compound (0.82 g;R_(f)=0.4, 29% yield over 3 steps) as an off-white foamy solid. MS m/z566.4 (M+H, expected 565.20 for C₃₀H₃₂ClN₃O₆).

Preparation 96 Biphenyl-2-ylcarbamic Acid1-[2-(2-Chloro-4-hydroxymethyl-5-methoxy-phenylcarbamoyl)ethyl]piperidin-4-ylEster

[0725] To a solution of the product of Preparation 95(0.82 mg, 1.45mmol) in a mixture of THF (4.5 mL) and methanol (0.5 mL) at 0° C. wasadded lithium borohydride (32 mg, 1.45 mmol). The reaction mixture wasallowed to warm to room temperature and was stirred for 41 h. Thereaction was then quenched by adding 1N aqueous hydrochloric acid at 0°C. until no more bubbling was observed and this mixture was stirred for10 min. The solvent was removed in vacuo and the residue was dissolvedin acetonitrile (about 2 mL). This solution was purified by prep-RP-HPLC(gradient: 2 to 50% acetonitrile in water with 0.05% TFA). Theappropriate fractions were collected and combined and lyophilized togive the title compound as a trifluoroacetate salt. This salt wastreated with isopropyl acetate (10 mL) and 1N aqueous sodium hydroxide(10 mL) and the organic layer was collected, dried (Na₂SO₄), filteredand the solvent was removed in vacuo to give the title compound (161 mg,21% yield) as a white foamy solid. MS m/z 538.4 (M+H, expected 537.20for C₂₉H₃₂ClN₃O₅).

Preparation 97 Biphenyl-2-ylcarbamic Acid1-[2-(2—Chloro-4-formyl-5-methoxyphenylcarbamoyl)-ethyl]piperidin-4-ylEster

[0726] To a solution of the product of Preparation 96 (161 mg, 0.3 mmol)in dichloromethane (3 mL) was added dimethyl sulfoxide (213 μL, 3.0mmol) and diisopropylethylamine (261 μL, 1.5 mmol). This mixture wascooled to −20° C. and sulfur trioxide pyridine complex (238 mg, 1.5mmol) was added slowly. After 30 min, the reaction mixture was quenchedby adding water (about 3 mL). The layers were separated and the organiclayer was dried (Na₂SO₄), filtered and the solvent was removed in vacuoto give the title compound as a light yellow solid. MS m/z 536.3 (M+H,expected 535.19 for C₂₉H₃₀ClN₃O₅).

Preparation 98 Biphenyl-2-ylcarbamic Acid1-[2-(4-{[(R)-2-(tert-Butyldimethylsilanyloxy)-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-2-chloro-5-methoxy-phenylcarbamoyl)ethyl]piperidin-4-ylEster

[0727] To the product from Preparation 97 in a mixture ofdichloromethane (0.5 mL) and methanol (0.5 mL) was added5-[(R)-2-amino-1-(tert-butyldimethylsilanyloxy)ethyl]-8-hydroxy-1H-quinolin-2-oneacetate (124.1 mg, 3.1 mmol) and the resulting mixture was stirred atroom temperature for 1.5 h. Sodium triacetoxyborohydride (190.7 mg, 0.9mmol) was added and the resulting mixture was stirred at roomtemperature for 15 h. The reaction was quenched by adding water (about0.2 mL) and the mixture was concentrated in vacuo to give the titlecompound, which was used without further purification. MS m/z 854.5(M+H, expected 853.36 for C₄₆H₅₆ClN₅O₇Si).

Example 25 Biphenyl-2-ylcarbamic Acid1-[2-(2-Chloro-4-{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-5-methoxyphenylcarbamoyl)ethyl]piperidin-4-ylEster Ditrifluoroacetate

[0728] To a suspension of the product of Preparation 98 indichloromethane (1.0 mL, 0.3 M) was added triethylamine trihydrofluoride(245 μL, 1.5 mmol). This mixture was stirred at room temperature for 45h and then the mixture was concentrated in vacuo. The residue wasdissolved in a mixture of DMF (0.5 mL), acetonitrile/water (1:1, with0.1% TFA, 0.6 mL), TFA (0.3 mL) and acetonitrile (about 1 mL) and thismixture was purified by prep-RP-HPLC (gradient: 2 to 50% acetonitrile inwater with 0.05% TFA). The appropriate 30 fractions were collected andcombined and lyophilized to give the title compound (100 mg, 34% yield,98.7% pure by HPLC) as an off-white solid. MS m/z 740.5 (M+H, expected739.28 for C₄₀H₄₂ClN₅O₇).

[0729] Using the methods described above and the appropriate startingmaterials, the following compounds were prepared. Ex. Compound MS 26Biphenyl-2-ylcarbamic acid 1-{7-[(R)-2-hydroxy- 613.52-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]heptyl}piperidin-4-yl ester 27 Biphenyl-2-ylcarbamic acid1-{8-[(R)-2-hydroxy- 627.5 2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]octyl}piperidin-4-yl ester 28 Biphenyl-2-ylcarbamic acid1-{2-[3-(4-{2- 705.3[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenyl)ureido]ethyl}piperidin- 4-yl ester 29Biphenyl-2-ylcarbamic acid 1-[3-(4-{2-[(R)- 682.42-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}piperidin-1-yl)-3- oxopropyl]piperidin-4-yl ester 30Biphenyl-2-ylcarbamic acid 1-{2-[(4-{[(R)-2-hydroxy- 682.72-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-cyclohexanecarbonyl)amino]ethyl}piperidin-4-yl ester 31Biphenyl-2-ylcarbamic acid 1-[2-({(1R,3S)-3-[(R)-2-(3- 630.2formylamino-4-hydroxyphenyl)-2-hydroxyethylamino]-cyclopentanecarbonyl}amino)ethyl]piperidin-4-yl ester 32Biphenyl-2-ylcarbamic acid 1-[2-(3-{5- 647.5[(R)-2-(3-formylamino-4-hydroxyphenyl)-2-hydroxyethylamino]pentyl}ureido)ethyl]piperidin- 4-yl ester 33Biphenyl-2-ylcarbamic acid 1-[2-(4-{2-[(R)-2-hydroxy- 662.52-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylamino)ethyl]piperidin- 4-yl ester 34Biphenyl-2-ylcarbamic acid 1-[3-(3-{5-[2- 661.3(3-formylamino-4-hydroxyphenyl)-2-hydroxyethylamino]pentyl}ureido)propyl]piperidin- 4-yl ester 35Biphenyl-2-ylcarbamic acid 1-{2-[(4-{2-[(R)-2-hydroxy-2-(8- 697.5hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}piperidine-1-carbonyl)amino]ethyl}piperidin-4-yl ester 36Biphenyl-2-ylcarbamic acid 1-[4-(4-{2-[(R)-2- 724.5hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}-phenylamino)benzyl]piperidin- 4-yl ester 37Biphenyl-2-ylcarbamic acid 1-[2-(3-{[(R)-2-hydroxy- 690.32-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-benzylcarbamoyl)ethyl]piperidin- 4-yl ester 383-[4-(3-Biphenyl-2-yl-ureido)piperidin-1-yl]-N-(4-{[ 675.5(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenyl)propionamide 39 Biphenyl-2-ylcarbamic acid1-{2-[(6-{[(R)-2- 691.5 hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-1)ethylamino]methyl}pyridin-2- ylmethyl)carbamoyl]ethyl}piperidin-4-ylester 40 Biphenyl-2-ylcarbamic acid 1-[2-(4-{[(R)-2- 682.7hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylarnino]methyl}- cyclohexylcarbamoyl)ethyl]piperidin-4-yl ester41 Biphenyl-2-ylcarbamic acid 1-[2-(4-{[(R)-2-hydroxy-2-(8- 682.7hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-cyclohexylcarbamoyl)ethyl]piperidin-4-yl ester 42 Biphenyl-2-ylcarbamicacid 1-[2-({(1R,3S)-3-[(R)-2- 654.8hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]-cyclopentanecarbonyl}amino)ethyl]piperidin- 4-yl ester 43Biphenyl-2-ylcarbamic acid 1-{2-[(3-{[(R)-2-hydroxy- 690.42-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-benzoyl)methylamino]ethyl} piperidin-4-yl ester 44 Biphenyl-2-ylcarbamicacid 1-{2-[(4-{[(R)-2-hydroxy-2-(8- 696.5hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-cyclohexanecarbonyl)methylamino]ethyl}piperidin-4-yl ester 45Biphenyl-2-ylcarbamic acid 1-[2-(4-{[2-hydroxy-2- NA(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenylcarbamoyl)ethyl]piperidin- 4-yl ester 46Biphenyl-2-ylcarbamic acid 1-[2-(4-{(S)-1-[(R)-2- 690.7hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylcarbamoyl)ethyl]piperidin- 4-yl ester 47Biphenyl-2-ylcarbamic acid 1-[2-(4-{(R)-1-[(R)-2- 690.7hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylcarbamoyl)ethyl]piperidin- 4-yl ester 48Biphenyl-2-ylcarbamic acid 1-((S)-1-{5-[(R)-2- 682.7hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]pentanoyl}pyrrolidin-2-ylmethyl)piperidin- 4-yl ester 49Biphenyl-2-ylcarbamic acid 1-[(S)-1-(4-{[(R)- 716.82-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-benzoyl)pyrrolidin-2- ylmethyl]piperidin-4-ylester 50 Biphenyl-2-ylcarbamic acid 1-[2-(4-{[(R)-2- 652.6(3-formylamino-4-hydroxy-phenyl)-2-hydroxy-ethylamino]methyl}phenylcarbamoyl)ethyl]piperidin- 4-yl ester 51Biphenyl-2-ylcarbamic acid 1-[2-(4-{(R)-1- 666.5[(R)-2-(3-formylamino-4-hydroxy-phenyl)-2-hydroxy-ethylamino]ethyl}phenylcarbamoyl)ethyl]piperidin- 4-yl ester 52Biphenyl-2-ylcarbamic acid 1-[2-(4-chloro-3- 710.5{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenylcarbamoyl)ethyl]piperidin- 4-yl ester 53N-{2-[4-(3-Biphenyl-2-yl-ureido)-piperidin-1-yl]ethyl}- 675.54-{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}benzamide 541-Biphenyl-2-yl-3-{1-[3-(4-{2-[(R)-2-hydroxy-2-(8-hydroxy- 681.72-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}piperidin-1-yl)-3-oxo-propyl]piperidin-4-yl}urea 553-[4-(3-Biphenyl-2-yl-ureido)piperidin-1-yl]-N-(3- 689.5{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-benzyl)propionamide 56 Biphenyl-2-ylcarbamicacid 1-(2-fluoro-3-{[(R)-2-hydroxy-2-(8- 637.5hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-benzyl)piperidin-4-yl ester 57 Biphenyl-2-ylcarbamic acid1-[2-(3-{[(R)-2-hydroxy-2-(8- 690.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-4-methyl-phenylcarbamoyl)ethyl]piperidin-4-yl ester 58Biphenyl-2-ylcarbamic acid 1-[2-(2-chloro-5- 710.6{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenylcarbamoyl)ethyl]piperidin- 4-yl ester 59Biphenyl-2-ylcarbamic acid 1-[2-(2,6-dichloro-4- 745.2{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenylcarbamoyl)ethyl]piperidin- 4-yl ester 60Biphenyl-2-ylcarbamic acid 1-[1-(4-{[(R)- 730.82-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}benzoyl)-piperidin-4- ylmethyl]piperidin-4-yl ester61 Biphenyl-2-ylcarbamic acid 1-[2-(4-{[(R)-2-(3- 652.5formylamino-4-hydroxy-phenyl)-2-hydroxyethylamino]methyl}-benzoylamino)ethyl]piperidin-4-yl ester 62 Biphenyl-2-ylcarbamic acid1-{2-[ethyl-(4- 704.5{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenyl)carbamoyl]ethyl}piperidin- 4-yl ester 63Biphenyl-2-ylcarbamic acid 1-(3-{4-[(R)-2-hydroxy-2- NA(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]piperidin-1-yl}-3-oxo-propyl)piperidin-4-yl ester 64 Biphenyl-2-ylcarbamic acid1-[2-(4-{2-[(R)-2- 690.3hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylcarbamoyl)ethyl]piperidin- 4-yl ester 65Biphenyl-2-ylcarbamic acid 1-{2-[(5-{[(R)-2-hydroxy-2-(8- 682.5hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-thiophene-2-carbonyl)amino]ethyl}piperidin-4-yl ester 66Biphenyl-2-ylcarbamic acid 1-{2-[(4-{[(R)-2-hydroxy-2-(8- 735.7hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-3-nitro-benzoyl)methylamino]ethyl} piperidin-4-yl ester 67Biphenyl-2-ylcarbamic acid 1-[2-(4-{[(R)-2-(3- 658.8formylamino-4-hydroxyphenyl)-2-hydroxyethylamino]methyl}-cyclohexylcarbamoyl)ethyl]piperidin-4-yl ester 68 Biphenyl-2-ylcarbamicacid 1-[2-({4-[(R)-2- 682.7hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]cyclohexanecarbonyl}- methylamino)ethyl]piperidin-4-ylester 69 Biphenyl-2-ylcarbamic acid 1-(2-fluoro-3-{4-[(R)-2- 720.5hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]piperidin-1-ylmethyl}benzyl)piperidin- 4-yl ester 70Biphenyl-2-ylcarbamic acid 1-{2-[(6-{[(R)-2- 677.5hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}pyridine-3- carbonyl)amino]ethyl}piperidin-4-ylester 71 Biphenyl-2-ylcarbamic acid 1-[3-(4-{[(R)-2- 654.5hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}piperidin-1-yl)-propyl]piperidin- 4-yl ester 72Biphenyl-2-ylcarbamic acid 1-[2-(4-{2-[(R)-2- 666.5(3-formylamino-4-hydroxyphenyl)-2-hydroxy-ethylamino]ethyl}phenylcarbamoyl)ethyl]piperidin- 4-yl ester 73Biphenyl-2-ylcarbamic acid 1-[2-(4-{2-[(R)-2- 690.3hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylamino)benzyl]piperidin- 4-yl ester 74Biphenyl-2-ylcarbamic acid 1-[2-fluoro-3-(4-{2- 748.5[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}piperidin-1- ylmethyl)benzyl]piperidin-4-yl ester75 Biphenyl-2-ylcarbamic acid 1-[3-(4-{2-[(tf)-2- 676.4hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylamino)propyl]piperidin-4-yl ester 76Biphenyl-2-ylcarbamic acid 1-[2-(3-chloro-4-{[(R)- 710.22-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenylcarbamoyl)ethyl]piperidin- 4-yl ester 77Biphenyl-2-ylcarbamic acid 1-[2-(4-{[(R)-2-hydroxy-2-(8- 769.2hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-2-trifluoromethoxy-phenylcarbamoyl)ethyl]piperidin-4-yl ester 78Biphenyl-2-ylcarbamic acid 1-{3-[3-(4-{2- 752.6[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylamino)phenyl]propyl}piperidin- 4-yl ester 79Biphenyl-2-ylcarbamic acid 1-[3-(4-{2-[(5)-2- NAhydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylamino)benzyl]piperidin- 4-yl ester 80Biphenyl-2-ylcarbamic acid 1-[2-(4-{[(R)-2-hydroxy-2-(8- 802.1hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-2-iodo-phenylcarbamoyl)ethyl]piperidin-4-yl ester 81Biphenyl-2-ylcarbamic acid 1-[2-(2-chloro-4- 724.2{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-6-methylphenylcarbamoyl)ethyl]piperidin-4-yl ester 82Biphenyl-2-ylcarbamic acid 1-(2-{5-[2-hydroxy-2- 656.5(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]pentylcarbamoyl}ethyl)-piperidin- 4-yl ester 83Biphenyl-2-ylcarbamic acid 1-[2-(2-bromo-4- 756.2{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenylcarbamoyl)ethyl]piperidin- 4-yl ester 84Biphenyl-2-ylcarbamic acid 1-{3-[2-(4-{2- 752.8[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylamino)- phenyl]propyl}piperidin-4-yl ester85 Biphenyl-2-ylcarbamic acid 1-[2-fluoro-3-(4-{3- 762.8[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]propyl}piperidin-1- ylmethyl)benzyl]piperidin-4-yl ester86 Biphenyl-2-ylcarbamic acid 1-[2-(4-{[(R)-2-hydroxy-2-(8- 706.3hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-2-methoxy-phenylcarbamoyl)ethyl]piperidin-4-yl ester 87Biphenyl-2-ylcarbamic acid 1-[5-(4-{2-[(R)-2- 704.3hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylamino)pentyl]piperidin-4-yl ester 88Biphenyl-2-ylcarbamic acid 1-{2-[1-(4-{2- 730.8[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenyl)-piperidin-4- yl]ethyl}piperidin-4-yl ester89 Biphenyl-2-ylcarbamic acid 1-[2-(4-{2-[(R)-2-hydroxy-2-(8- 704.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]-1-methyl-ethyl}phenylcarbamoyl)ethyl]piperidin-4-yl ester 90Biphenyl-2-ylcarbamic acid 1-{2-[4-(4-{2- 744.4[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylamino)cyclohexyl]ethyl}piperidin- 4-yl ester91 Biphenyl-2-ylcarbamic acid 1-[2-(2-fluoro-4- 694.3{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phehylcarbamoyl)ethyl]piperidin- 4-yl ester 92Biphenyl-2-ylcarbamic acid 1-{2-[3-(4-{2- 738.8[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylamino)phenyl]ethyl}piperidin- 4-yl ester 93Biphenyl-2-ylcarbamic acid 1-[2-(2,5-difluoro-4- 712.3{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenylcarbamoyl)ethyl]piperidin- 4-yl ester 94Biphenyl-2-ylcarbamic acid 1-[2-(4-{2-[(R)-2-hydroxy-2- 690.3(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}-benzoylamino)ethyl]piperidin-4-yl ester 95 Biphenyl-2-ylcarbamic acid1-[6-(4-{[(R)-2- 717.5 hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}piperidin-1-ylmethyl)pyridin-2-ylmethyl]piperidin-4-yl ester 96 Biphenyl-2-ylcarbamic acid1-[2-(4-{2-[(R)-2-hydroxy-2-(8- 740.6hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}-naphthalen-1-ylcarbamoyl)ethyl]piperidin-4-yl ester 97Biphenyl-2-ylcarbamic acid 1-{2-[1-(4-{[(R)-2-hydroxy- 744.42-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-benzoyl)piperidin-4-yl]ethyl}piperidin-4-yl ester 98Biphenyl-2-ylcarbamic acid 1-[3-(4-{3-[(R)-2- 704.2hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]propionylamino}phenyl)propyl]piperidin- 4-yl ester 99Biphenyl-2-ylcarbamic acid 1-[3-(4-{[(R)-2- 663.7hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)-propyl]piperidin- 4-yl ester 100Biphenyl-2-ylcarbamic acid 1-[2-(5-{[(R)-2-hydroxy-2-(8- 673.7hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-1H-benzoimidazol-2-yl)ethyl]piperidin-4-yl ester 101Biphenyl-2-ylcarbamic acid 1-[2-(4-{3-[(R)-2- 696.4hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]propionylamino}cyclohexyl)ethyl]piperidin- 4-yl ester102 Biphenyl-2-ylcarbamic acid 1-[2-(4-{5-[(R)- 724.42-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]pentanoylamino}cyclohexyl)ethyl]piperidin- 4-yl ester103 Biphenyl-2-ylcarbamic acid 1-[2-(4-{6-[(R)-2-hydroxy- 738.42-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]-hexanoylamino}cyclohexyl)ethyl]piperidin-4-yl ester 104Biphenyl-2-ylcarbamic acid 1-[2-(1-{3-[(R)-2- 682.4hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]propionyl}piperidin-4-yl)ethyl]piperidin- 4-yl ester 105Biphenyl-2-ylcarbamic acid 1-{2-[3-(4- 691.7{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenyl)ureido]ethyl}piperidin- 4-yl ester 106Biphenyl-2-ylcarbamic acid 1-{2-[(2-{4-[(R)-2-hydroxy-2-(8- 682.7hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]cyclohexyl}-ethyl)methylamino]ethyl}piperidin-4-yl ester 107 Biphenyl-2-ylcarbamicacid 1-[2-(2,3,5,6-tetrafluoro- 748.24-{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenylcarbamoyl)ethyl]piperidin- 4-yl ester 108Biphenyl-2-ylcarbamic acid 1-[2-(4-{[(R)-2-hydroxy-2-(8- 928.0hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-2,6-diiodo-phenylcarbamoyl)ethyl]piperidin-4-yl ester 109Biphenyl-2-ylcarbamic acid 1-[2-(1-{4-[(R)-2-hydroxy-2- 696.4(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]-butyryl}piperidin-4-yl)ethyl]piperidin-4-yl ester 110Biphenyl-2-ylcarbamic acid 1-[2-(1-{5-[(R)-2- 710.4hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]pentanoyl}piperidin-4-yl)ethyl]piperidin- 4-yl ester 111Biphenyl-2-ylcarbamic acid 1-[2-(1-{6-[(R)-2-hydroxy- 724.42-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]-hexanoyl}piperidin-4-yl)ethyl]piperidin-4-yl ester 112Biphenyl-2-ylcarbamic acid 1-[2-(4-{[(R)-2-hydroxy-2- 690.5(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-benzylcarbamoyl)ethyl]piperidin-4-yl ester 113 Biphenyl-2-ylcarbamicacid 1-[2-(4-{[(R)-2-(3- 666.5formylamino-4-hydroxy-phenyl)-2-hydroxyethylamino]methyl}-benzylcarbamoyl)ethyl]piperidin-4-yl ester 114 Biphenyl-2-ylcarbamicacid 1-{2-[3-(4-{[(R)-2-hydroxy-2-(8- 705.6hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-benzyl)ureido]ethyl}piperidin-4-yl ester 115 Biphenyl-2-ylcarbamic acid1-{2-[3-(4- 681.7 {[(R)-2-(3-formylamino-4-hydroxyphenyl)-2-hydroxyethylamino]methyl}benzyl)- ureido]ethyl}piperidin-4-yl ester 116Biphenyl-2-ylcarbamic acid 1-[2-(4-{[(R)-2-hydroxy-2-(8- 690.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-2-methyl-phenylcarbamoyl)ethyl]piperidin-4-yl ester 117Biphenyl-2-ylcarbamic acid 1-(3-{4-[2-(4- 774.4{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}piperidin-1-yl)ethyl]phenoxy}propyl)-piperidin-4-yl ester 118Biphenyl-2-ylcarbamic acid 1-[2-(3-{2-[(R)-2-hydroxy-2- 690.4(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}-benzylcarbamoyl)ethyl]piperidin-4-yl ester 119 Biphenyl-2-ylcarbamicacid 1-[2-(3-{[(R)-2- 649.5hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)ethyl]piperidin- 4-yl ester 120Biphenyl-2-ylcarbamic acid 1-(2-{[2-(4- 720.4{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)acetyl]methylamino}ethyl)- piperidin-4-ylester 121 Biphenyl-2-ylcarbamic acid 1-(2-{[2-(3- 720.4{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)acetyl]methylamino}ethyl)- piperidin-4-ylester 122 Biphenyl-2-ylcarbamic acid 1-{2-[(5-{[(R)-2- 680.3hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}furan-2-carbonyl)methylamino]ethyl}piperidin-4-yl ester 123Biphenyl-2-ylcarbamic acid 1-{2-[(5-{[(R)-2-hydroxy-2- 696.2(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-thiophene-2-carbonyl)methylamino]ethyl}piperidin-4-yl ester 124Biphenyl-2-ylcarbamic acid 1-[2-(4-{2-[(R)- 679.32-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethoxy}phenoxy)ethyl]piperidin-4-yl ester 125Biphenyl-2-ylcarbamic acid 1-{2-[4-(4-{[(R)-2-hydroxy- 758.42-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-benzoylamino)cyclohexyl]ethyl}piperidin-4-yl ester 126Biphenyl-2-ylcarbamic acid 1-(2-{4-[2-(2- 788.4{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)acetylamino]cyclo-hexyl}ethyl)-piperidin-4-yl ester 127 Biphenyl-2-ylcarbamic acid1-(2-{4-[2-(3-{[(R)- 788.42-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)acetylamino]cyclo-hexyl}ethyl)-piperidin-4-yl ester 128 Biphenyl-2-ylcarbamic acid1-(2-{4-[2-(4-{[(R)- 788.42-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)acetylamino]cyclo-hexyl}ethyl)-piperidin-4-yl ester 129 Biphenyl-2-ylcarbamic acid1-(2-{4-[(5-{[(R)-2- 748.4hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}furan-2-carbonyl)amino]cyclohexyl}ethyl)piperidin-4-yl ester 130Biphenyl-2-ylcarbamic acid 1-(2-{4-[(5- 764.4{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-thiophene-2-carbonyl)amino]cyclohexyl}ethyl)piperidin-4-yl ester 131Biphenyl-2-ylcarbamic acid 1-(2-{1-[2-(2- 774.4{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)acetyl]piperidin-4-yl}ethyl)-piperidin-4-yl ester 132 Biphenyl-2-ylcarbamic acid1-(2-{1-[2-(3- 774.4{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)acetyl]piperidin-4-yl}ethyl)-piperidin-4-yl ester 133 Biphenyl-2-ylcarbamic acid1-(2-{1-[2-(4- 774.4{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)acetyl]piperidin-4-yl}ethyl)-piperidin-4-yl ester 134 Biphenyl-2-ylcarbamic acid1-{2-[1-(5- 734.4{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}furan-2-carbonyl) piperidin-4-yl]ethyl}piperidin-4-yl ester 135 Biphenyl-2-ylcarbamic acid1-{2-[1-(5- 750.2{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-thiophene-2-carbonyl)piperidin-4-yl]ethyl}piperidin-4-yl ester 136 Biphenyl-2-ylcarbamic acid1-{2-[4-(3- 752.4 {[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-benzoylamino)phenyl]ethyl}piperidin-4-yl ester 137 Biphenyl-2-ylcarbamicacid 1-{2-[4-(4- 752.4 {[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-benzoylamino)phenyl]ethyl}piperidin-4-yl ester 138 Biphenyl-2-ylcarbamicacid 1-(2-{4-[2-(2- 782.4 {[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)acetyl-amino]phenyl}ethyl)-piperidin-4-yl ester 139 Biphenyl-2-ylcarbamic acid1-(2-{4-[2-(3- 782.4{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl} phenoxy)acetyl-amino]phenyl}ethyl)-piperidin-4-yl ester 140 Biphenyl-2-ylcarbamic acid1-(2-{4-[2-(4- 782.4{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)acetyl-amino]phenyl}ethyl)-piperidin-4-yl ester 141 Biphenyl-2-ylcarbamic acid1-(2-{4-[(5- 742.4{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}furan-2-carbonyl)amino]phenyl}ethyl)piperidin-4-yl ester 142Biphenyl-2-ylcarbamic acid 1-(2-{4-[(5- 758.2{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-thiophene-2-carbonyl)amino]phenyl}ethyl)piperidin-4-yl ester 143Biphenyl-2-ylcarbamic acid 1-{2-[4-(3- 758.4{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-benzoylamino)cyclo- hexyl]ethyl}piperidin-4-ylester 144 Biphenyl-2-ylcarbamic acid 1-[3-(3- 663.4{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)-propyl]piperidin- 4-yl ester 145Biphenyl-2-ylcarbamic acid 1-[2-hydroxy-3-(4-{2-[(R)- 692.32-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylamino)propyl]piperidin- 4-yl ester 146Biphenyl-2-ylcarbamic acid 1-[4-(4-{2-[(R)- 690.42-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylamino)butyl]piperidin- 4-yl ester 147Biphenyl-2-ylcarbamic acid 1-{2-[4-({2-[(R)-2- 733.3hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]acetylamino}-methyl)phenyl-carbamoyl]ethyl}piperidin-4-yl ester 148 Biphenyl-2-ylcarbamic acid1-{2-[4-(2-{2- 747.4[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]acetylamino}-ethyl)phenyl-carbamoyl]ethyl}piperidin-4-yl ester 149 Biphenyl-2-ylcarbamic acid1-{2-[(4- 696.6 {[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-cyclohexyl-methyl)carbamoyl]ethyl}piperidin-4-yl ester 150 Biphenyl-2-ylcarbamicacid 1-(2-{6-[(R)-2-hydroxy- 656.62-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]hexanoylamino}-ethyl)piperidin- 4-yl ester 151Biphenyl-2-ylcarbamic acid 1-[2-(3-{2-[(R)- 679.32-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethoxy}phenoxy)ethyl]piperidin- 4-yl ester 152Biphenyl-2-ylcarbamic acid 1-[2-(2-{2-[(S)- 679.32-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethoxy} phenoxy)ethyl]piperidin- 4-yl ester 153Biphenyl-2-ylcarbamic acid 1-[2-(2-{[(R)- 711.32-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)benzyl]piperidin- 4-yl ester 154Biphenyl-2-ylcarbamic acid 1-(2-{6-[(R)-2- 670.4hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]hexylcarbamoyl}ethyl)piperidin- 4-yl ester 155Biphenyl-2-ylcarbamic acid 1-[2-({(1R,35)-3- 654.8[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]cyclopentanecarbonyl}amino)ethyl]piperidin- 4-yl ester 156Biphenyl-2-ylcarbamic acid 1-[3-(4-{3-[(R)- 675.52-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]propyl}phenyl)propyl]piperidin- 4-yl ester 157Biphenyl-2-ylcarbamic acid 1-[3-(4-{2-[(R)- 661.32-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenyl)propyl]piperidin- 4-yl ester 158Biphenyl-2-ylcarbamic acid 1-[4-(4-{2-[(R)- 675.52-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenyl)butyl]piperidin- 4-yl ester 159Biphenyl-2-ylcarbamic acid 1-[3-(5-{3-[(R)-2- 665.6hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5- yl)ethylamino]propyl}furan-2-yl)propyl]piperidin- 4-yl ester 160 Biphenyl-2-ylcarbamic acid1-{2-[3-(4-{2- 719.2[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenyl)-1- methylureido]ethyl}piperidin-4-yl ester161 Biphenyl-2-ylcarbamic acid 1-{2-[l-(4-{2- 773.3[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylcarbamoyl)piperidin-4-yl]ethyl}piperidin-4-yl ester 162 Biphenyl-2-ylcarbamic acid1-[3-(3-{3-[(R)- 675.52-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]propyl}phenyl)propyl]piperidin- 4-yl ester 163Biphenyl-2-ylcarbamic acid 1-[3-(5-{3- 669.6[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]propyl}tetrahydrofuran-2- yl)propyl]piperidin-4-yl ester164 Biphenyl-2-ylcarbamic acid 1-[2-(4-{2-[(R)-2-hydroxy-2-(8- 706.5hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethylcarbamoyl}phenoxy)ethyl]piperidin-4-yl ester 165(5-Bromobiphenyl-2-yl)carbamic acid 1-{9-[2- NAhydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]nonyl}-piperidin-4-yl ester 166(2′-Fluorobiphenyl-2-yl)carbamic acid 1-{9- 659.5[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]nonyl}-piperidin-4-yl ester 167(3′-Chloro-3,5-difluorobiphenyl-2-yl)carbamic acid 1- 711.8{9-[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)-ethylamino]nonyl}piperidin- 4-yl ester 168(3′,5′-Dichloro-3,5-difluorobiphenyl- 745.5 2-yl)carbamic acid1-{9-[(R)-2-hydroxy-2- (8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)-ethylamino]nonyl}piperidin-4-yl ester 169(3,5-Difluorobiphenyl-2-yl)carbamic acid 1-{9- 677.5[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]nonyl}-piperidin-4-yl ester

Preparation 99 Biphenyl-2-ylcarbamic Acid1-[2-(4-[1,3]dioxolan-2-ylphenylcarbamoyl)-ethyl]-4-methylpiperidin-4-ylEster

[0730] A mixture of biphenyl-2-ylcarbamic acid 4-methylpiperidin-4-ylester (2.73 g, 8.79 mmol) and N-(4-[1,3]dioxolan-2-yl-phenyl)acrylamide(2.05 g, 8.80 mmol) were heated in 100 mL of 1:1methanol/dichloromethane at 50° C. under nitrogen for 1 h. The solutionwas then diluted with ethyl acetate and the organic layer was washedwith water, brine, dried (MgSO₄) and concentrated under reduced pressureto give the title compound. MS m/z calcd for C₃₁H₃₅N₃O₅ (M+H)⁺ 530.6;found 530.4.

Preparation 100 Biphenyl-2-ylcarbamic Acid1-[2-(4-Formylphenylcarbamoyl)ethyl]-4-methylpiperidin-4-yl Ester

[0731] The product of Preparation 99 was redissolved in 40 mL ofmethanol and 25 mL of aqueous 1 N hydrochloric acid was added. Theresulting mixture was stirred at room temperature overnight and theorganic solvent was removed under reduced pressure. The residue wasdissolved in ethyl acetate and the organic layer was washed with water,brine, dried (MgSO₄) and the solvent removed under reduced pressure. Theproduct was triturated with dichloromethane to give the title compoundas a white powder (2.47 g). LCMS (2-90) R_(t)=4.27 min; MS m/z calcd forC₂₉H₃₁N₃O₄ (M+H)⁺ 486.6, found 486.5.

Preparation 101 Biphenyl-2-ylcarbamic Acid1-[2-(4-{[(R)-2-(tert-butyldimethylsilanyloxy)-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenylcarbamoyl)ethyl]-4-methyl-piperidin-4-ylEster

[0732] A mixture of the product of Preparation 100 (1.70 g, 3.51 mmol)and5-[(R)-2-amino-1-(tert-butyldimethylsilanyloxy)ethyl]-8-hydroxy-1H-quinolin-2-oneacetate (1.65 g, 4.19 mmol) in 40 mL of 1:1 methanol and dichloromethanewas stirred at room temperature overnight. Sodium triacetoxyborohydride(2.23 g, 10.5 mmol) was then added in one portion and the reactionmixture was stirred at room temperature for 6 hr. The reaction was thenquenched with water and diluted with ethyl acetate. The layers wereseparated and the organic layer was washed with saturated sodiumbicarbonate, brine, dried (MgSO₄) and the solvent removed under reducedpressure to give the title compound (2.9 g). MS m/z calcd forC₄₆H₅₇N₅O₆Si (M+H)+ 805.0, found 804.6.

Example 170 Biphenyl-2-ylcarbamic Acid1-[2-(4-{[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-phenylcarbamoyl)ethyl]-4-methylpiperidin-4-ylEster

[0733] The product of Preparation 101 (2.9 g, 3.6 mmol) was dissolved in75 mL of dichloromethane and triethylamine trihydrofluoride (0.85 mL,5.2 mmol) was added. The resulting mixture was stirred at roomtemperature overnight and then the solvent was removed under reducedpressure to give the crude product as an oil. The product was thendissolved in acetic acid/water (1:1) and purified by prep HPLC to givethe title compound as an off-white solid. LCMS (2-90) R_(t)=3.67 min.;MS m/z calcd C₄₀H₄₃N₅O₆ (M+H)⁺ 690.8, found 690.3.

[0734] Using the methods described herein and the appropriate startingmaterials, the following compounds can be prepared. Ex. Compound MS 171Biphenyl-2-ylcarbamic acid 1-[2-(4-{[(R)-2-(3- NAformylamino-4-hydroxyphenyl)-2-hydroxyethyl-amino]methyl}phenylcarbamoyl)-ethyl]-4-methyl- piperidin-4-yl ester 172Biphenyl-2-ylcarbamic acid 1-{9-[(R)-2-hydroxy-2- NA(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl-amino]nonyl}-4-ethylpiperidin-4-yl ester 173 Biphenyl-2-ylcarbamic acid1-{9-[(R)-2-(3- NA formylamino-4-hydroxy-phenyl)-2-hydroxyethyl-amino]nonyl}-4-methylpiperidin-4-yl ester 174 Biphenyl-2-ylcarbamic acid1-(2-{5-[(R)-2-hydroxy-2- NA (8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]pentylcarbamoyl}-ethyl)-4- methylpiperidin-4-yl ester 175Biphenyl-2-ylcarbamic acid 1-(2-{5-[(R)-2- NA(3-formylamino-4-hydroxyphenyl)-2-hydroxyethyl-amino]pentylcarbamoyl}ethyl)-4- methylpiperidin-4-yl ester 176Biphenyl-2-ylcarbamic acid 1-(2-{6-[(R)-2- NAhydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]hexanoylamino}ethyl)- 4-methylpiperidin-4-yl ester 177Biphenyl-2-ylcarbamic acid 1-(2-{6-[(R)-2- NA(3-formylamino-4-hydroxyphenyl)-2-hydroxyethyl-amino]hexanoylamino}ethyl)-4-methylpiperidin- 4-yl ester 178Biphenyl-2-ylcarbamic acid 1-[2-(4-{[(R)-2- NAhydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-benzoylamino)ethyl]- 4-methylpiperidin-4-yl ester179 Biphenyl-2-ylcarbamic acid 1-[2-(4-{[(R)-2-(3- NAformylamino-4-hydroxyphenyl)-2-hydroxyethyl-amino]methyl}benzoylamino)-ethyl]-4- methylpiperidin-4-yl ester 180Biphenyl-2-ylcarbamic acid 1-{3-[4-(4-{2- 776.5[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylamino)phenyl]propyl}- 4-methylpiperidin-4-ylester 181 Biphenyl-2-ylcarbamic acid 1-[2-(2-chloro-4- 724.5{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenylcarbamoyl)ethyl]- 4-methylpiperidin-4-ylester 182 Biphenyl-2-ylcarbamic acid 1-[2-(2-chloro-4- 754.5{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-5-methoxyphenylcarbamoyl)ethyl]-4-methylpiperidin-4-yl ester

Preparation 102 Biphenyl-2-ylcarbamic acid(R)-(1-azabicyclo[3.2.1]oct-4-yl) Ester

[0735] 2-Biphenyl isocyanate (1.00 g, 5.12 mmol) and(R)-(−)-3-quinuclidinol hydrochloride (921 mg, 5.63 mmol) were heatedtogether in N,N-dimethylformamide (2.06 mL) at 110° C. for 12 h. Thereaction mixture was cooled and diluted with ethyl acetate (15 mL) andthen washed with saturated aqueous sodium bicarbonate (2×10 mL). Theorganic layer was extracted with 1 M hydrochloric acid (3×20 mL) and thecombined aqueous extracts were made basic to pH 8-9 with potassiumcarbonate. The aqueous layer was then extracted with ethyl acetate (3×20mL) and the combined organic layers were dried (magnesium sulfate) andsolvent was removed under reduced pressure to give the title compound asa yellow oil (1.64 g, 99% yield).

Preparation 103(R)-4-(Biphenyl-2-ylcarbamoyloxy)-1-(9-bromononyl)-1-azoniabicyclo[3.2.1]octaneBromide

[0736] To a stirred solution of the product of Preparation 102 (1.21 g,3.76 mmol) and triethylamine (1.05 mL, 7.52 mmol) in acetonitrile (18.8mL) was added 1,9-dibromononane (994 μL, 4.89 mmol) and the reactionmixture was heated at 50° C. for 4 h. The reaction mixture was thencooled and the solvent was removed under reduced pressure. The residuewas dissolved in dichloromethane (20 mL) and the organic layer waswashed with saturated aqueous sodium bicarbonate (10 mL), dried(magnesium sulfate) and solvent removed under reduced pressure. Thecrude product was purified by flash chromatography (10%methanol/dichloromethane, 0.5% ammonium hydroxide) to give the titlecompound (1.04 g, 1.97 mmol, 52% yield).

Preparation 104(R)-1-(9-N,N-Di(tert-butoxycarbonyl)aminononyl)-4-(biphenyl-2-ylcarbamoyloxy)-1-azoniabicyclo[3.2.1]octaneBromide

[0737] To a stirred solution of sodium hydride (60% dispersion inmineral oil) (126 mg, 3.15 mmol) in N,N-dimethylformamide (10 mL) underan atmosphere of nitrogen at 0° C., was added di-tert-butyliminodicarboxylate (513 mg, 2.36 mmol) in N,N-dimethylformamide (5 mL).The reaction mixture was stirred at room temperature for 15 min and thenit was cooled to 0° C. and the product of Preparation 103 (1.04 g, 1.97mmol) in N,N-dimethylformamide (5 mL) was added. The reaction mixturewas allowed to warm to room temperature over a 12 h period and then thesolvent was removed under reduced pressure to give the title compound,which was used without further purification.

Preparation 105(R)-1-(9-Aminononyl)-4-(biphenyl-2-ylcarbamoyloxy)-1-azoniabicyclo[3.2.1]octaneBromide

[0738] The product of Preparation 104 (1.31 g, 1.97 mmol) was dissolvedin dichloromethane (15 mL) and trifluoroacetic acid (5 mL) was addedslowly. The reaction mixture was stirred at room temperature for 1 h andthen the solvent was removed under reduced pressure. The residue wasdissolved in dichloromethane (20 mL) and washed with aqueous 1 M sodiumhydroxide (20 mL). The organic layer was extracted with 1 M hydrochloricacid (3×20 mL) and the combined aqueous extracts were made basic withpotassium carbonate and extracted with dichloromethane (3×20 mL). Thecombined organic layers were dried (magnesium sulfate) and solvent wasremoved under reduced pressure to give the title compound (210 mg, 23%yield over 2 steps).

Preparation 106(R)-1-{9-[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethylamino]-nonyl}-4-(biphenyl-2-ylcarbamoyloxy)-1-azoniabicyclo[3.2.1]octane Bromide

[0739] The product of Preparation 105 (210 mg, 0.45 mmol) and sodiumtriacetoxyborohydride (286 mg, 1.35 mmol) were stirred in dichloroethane(4.5 mL) at room temperature for 2 h and then the product of Preparation6 (163 mg, 0.50 mmol) was added. The reaction mixture was stirred for 12h and then diluted with dichloromethane (10 mL) and washed withsaturated aqueous sodium bicarbonate (10 mL), dried (magnesium sulfate)and solvent removed under reduced pressure. The crude reaction productwas purified by flash chromatography (10-50% methanol/dichloromethane,0.5% ammonium hydroxide) to give the title compound (131 mg, 38% yield).

Example 1834-(Biphenyl-2-ylcarbamoyloxy)-1-{9-[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)-ethylamino]nonyl}-1-azoniabicyclo[2.2.2]octaneBromide Ditrifluoroacetate

[0740] To a stirred solution of the product of Preparation 105 (131 mg,0.17 mmol) in methanol (1.8 mL) was added palladium (10 wt. % dry basison activated carbon; 39 mg) and the reaction mixture was placed under anatmosphere of hydrogen. After stirring for 12 h, the reaction mixturewas filtered through a pad of Celite, washing with methanol (2 mL) andsolvent removed under reduced pressure. The resulting residue waspurified by preparative HPLC to give the title compound as itsditrifluoroacetate salt (8 mg). MS m/z 667.5.

[0741] Using the methods described herein and the appropriate startingmaterials, the following compounds can be prepared. Ex. Compound MS 184Biphenyl-2-ylcarbamic acid 8-{9-[(R)-2-hydroxy-2-(8-hydroxy-2- 667.3oxo-1,2-dihydroquinolin-5-yl)ethylamino]nonyl}-8-aza-bicyclo[3.2.1]oct-3-yl ester 1857-(Biphenyl-2-ylcarbamoyloxy)-9-{9-[(R)-2-hydroxy-2-(8-hydroxy- 695.52-oxo-1,2-dihydroquinolin-5-yl)-ethylamino]nonyl}-9-methyl-3-oxa-9-azoniatricyclo[3.3.1.0*2,4*]nonane bromide 186Biphenyl-2-ylcarbamic acid 9-{9-[(R)-2-hydroxy-2-(8-hydroxy-2- 681.5oxo-1,2-dihydroquinolin-5-yl)ethylamino]nonyl}-3-oxa-9-aza-tricyclo[3.3.1.0*2,4*]non-7-yl ester

[0742] Members of the library of compounds prepared by the aboveprocedures were evaluated using the following in vitro and in vivoassays:

Preparation A Cell Culture and Membrane Preparation From CellsExpressing Human β₁, β₂ or β₃ Adrenergic Receptors

[0743] Chinese hamster ovarian (CHO) cell lines stably expressing clonedhuman β₁, β₂ or β₃ adrenergic receptors, respectively, were grown tonear confluency in Hams F-12 media with 10% FBS in the presence of 500μg/mL Geneticin. The cell monolayer was lifted with 2 mM EDTA in PBS.Cells were pelleted by centrifugation at 1,000 rpm, and cell pelletswere either stored frozen at −80° C. or membranes were preparedimmediately for use. For preparation of β₁ and β₂ receptor expressingmembranes, cell pellets were resuspended in lysis buffer (10 mMHEPES/HCl, 10 mM EDTA, pH 7.4 at 4° C.) and homogenized using atight-fitting Dounce glass homogenizer (30 strokes) on ice. For the moreprotease-sensitive β₃ receptor expressing membranes, cell pellets werehomogenated in lysis buffer (10 mM Tris/HCl, pH 7.4) supplemented withone tablet of “Complete Protease Inhibitor Cocktail Tablets with 2 mMEDTA” per 50 mL buffer (Roche Catalog No. 1697498, Roche MolecularBiochemicals, Indianapolis, Ind.). The homogenate was centrifuged at20,000×g, and the resulting pellet was washed once with lysis buffer byresuspension and centrifugation as above. The final pellet was thenre-suspended in ice-cold binding assay buffer (75 mM Tris/HCl pH 7.4,12.5 mM MgCl₂, 1 mM EDTA). The protein concentration of the membranesuspension was determined by the methods described in Lowry et al.,1951, Journal of Biological Chemistry, 193, 265; and Bradford,Analytical Biochemistry, 1976, 72, 248-54. All membranes were storedfrozen in aliquots at −80° C. or used immediately.

Preparation B Cell Culture and Membrane Preparation From CellsExpressing Human M₁, M₂, M₃ and M₄ Muscarinic Receptors

[0744] CHO cell lines stably expressing cloned human hM₁, hM₂, hM₃ andhM₄ muscarinic receptor subtypes, respectively, were grown to nearconfluency in HAM's F-12 media supplemented with 10% FBS and 250 μg/mLGeneticin. The cells were grown in a 5% CO₂, 37° C. incubator and liftedwith 2 mM EDTA in dPBS. Cells were collected by 5 minute centrifugationat 650×g, and cell pellets were either stored frozen at −80° C. ormembranes were prepared immediately for use. For membrane preparation,cell pellets were resuspended in lysis buffer and homogenized with aPolytron PT-2100 tissue disrupter (Kinematica AG; 20 seconds×2 bursts).Crude membranes were centrifuged at 40,000×g for 15 minutes at 4° C. Themembrane pellet was then resuspended with resuspension buffer andhomogenized again with the Polytron tissue disrupter. The proteinconcentration of the membrane suspension was determined by the methoddescribed in Lowry et al., 1951, Journal of Biochemistry, 193, 265. Allmembranes were stored frozen in aliquots at −80° C. or used immediately.Aliquots of prepared hM₅ receptor membranes were purchased directly fromPerkin Elmer and stored at −80° C. until use.

Assay Test Procedure A Radioligand Binding Assay for Human β₁, β₂ and β₃Adrenergic Receptors

[0745] Binding assays were performed in 96-well microtiter plates in atotal assay volume of 100 μL with 10-15 μg of membrane proteincontaining the human β₁, β₂ or β₃ adrenergic receptors in assay buffer(75 mM Tris/HCl pH 7.4 at 25° C., 12.5 mM MgCl₂, 1 mM EDTA, 0.2% BSA).Saturation binding studies for determination of K_(d) values of theradioligand were done using [³H]-dihydroalprenolol (NET-720, 100Ci/mmol, PerkinElmer Life Sciences Inc., Boston, Mass.) for the β₁ andβ₂ receptors and [¹²⁵I]-(−)-iodocyanopindolol (NEX-189, 220 Ci/mmol,PerkinElmer Life Sciences Inc., Boston, Mass.) at 10 or 11 differentconcentrations ranging from 0.01 nM to 20 nM. Displacement assays fordetermination of K_(i) values of test compounds were done with[³H]-dihydroalprenolol at 1 nM and [¹²⁵I]-(−)-iodocyanopindolol at 0.5nM for 10 or 11 different concentrations of test compound ranging from10 pM to 10 μM. Non-specific binding was determined in the presence of10 μM propranolol. Assays were incubated for 1 hour at 37° C., and thenbinding reactions were terminated by rapid filtration over GF/B for theβ₁ and β₂ receptors or GF/C glass fiber filter plates for the β₃receptors (Packard BioScience Co., Meriden, Conn.) presoaked in 0.3%polyethyleneimine. Filter plates were washed three times with filtrationbuffer (75 mM Tris/HCl pH 7.4 at 4° C., 12.5 mM MgCl₂, 1 mM EDTA) toremove unbound radioactivity. The plates were then dried and 50 μL ofMicroscint-20 liquid scintillation fluid (Packard BioScience Co.,Meriden, Conn.) was added and plates were counted in a Packard Topcountliquid scintillation counter (Packard BioScience Co., Meriden, Conn.).Binding data were analyzed by nonlinear regression analysis with theGraphPad Prism Software package (GraphPad Software, Inc., San Diego,Calif.) using the 3-parameter model for one-site competition. The curveminimum was fixed to the value for nonspecific binding, as determined inthe presence of 10 μM propranolol. K_(i) values for test compounds werecalculated from observed IC₅₀ values and the K_(d) value of theradioligand using the Cheng-Prusoff equation (Cheng Y, and Prusoff W H.,Biochemical Pharmacology, 1973, 22, 23, 3099-108).

[0746] In this assay, a lower K_(i) value indicates that a test compoundhas a higher binding affinity for the receptor tested. Exemplifiedcompound that were tested in this assay typically were found to have aK_(i) value of less than about 300 nM for the β₂ adrenergic receptor.For example, the compounds of Examples 3 and 6 were identified as havingK_(i) values of less than 10 nM.

[0747] If desired, the receptor subtype selectivity for a test compoundcan be calculated as the ratio of K_(i)(β₁)/K_(i)(β₂) orK_(i)(β₃)/K_(i)(β₂). Typically, compounds of this invention demonstratedgreater binding at the β₂ adrenergic receptor compared to the β₁ or β₃adrenergic receptor, i.e. K_(i)(β₁) or K_(i)(β₃) is typically greaterthan K_(i)(β₂). Generally, compounds having selectivity for the β₂adrenergic receptor over the β₁ or β₃ adrenergic receptors arepreferred; especially compounds having a selectivity greater than about5; and in particular, greater than about 8. By way of example, thecompounds of Examples 3 and 6 had a ratio of K_(i)(β₁)/K_(i)(β₂) greaterthan 8.

Assay Test Procedure B Radioligand Binding Assay for MuscarinicReceptors

[0748] Radioligand binding assays for cloned human muscarinic receptorswere performed in 96-well microtiter plates in a total assay volume of100 μL. CHO cell membranes stably expressing either the hM₁, hM₂, hM₃,hM₄ or hM₅ muscarinic subtype were diluted in assay buffer to thefollowing specific target protein concentrations (μg/well): 10 μg forhM₁, 10-15 μg for hM₂, 10-20 μg for hM₃, 10-20 μg for hM₄, and 10-12 μgfor hM₅ to get similar signals (cpm). The membranes were brieflyhomogenized using a Polytron tissue disrupter (10 seconds) prior toassay plate addition. Saturation binding studies for determining K_(D)values of the radioligand were performed usingL-[N-methyl-³H]scopolamine methyl chloride ([³H]-NMS) (TRK666, 84.0Ci/mmol, Amersham Pharmacia Biotech, Buckinghamshire, England) atconcentrations ranging from 0.001 nM to 20 nM. Displacement assays fordetermination of K_(i) values of test compounds were performed with[³H]-NMS at 1 nM and eleven different test compound concentrations. Thetest compounds were initially dissolved to a concentration of 400 μM indilution buffer and then serially diluted 5× with dilution buffer tofinal concentrations ranging from 10 pM to 100 μM. The addition orderand volumes to the assay plates were as follows: 25 μL radioligand, 25μL diluted test compound, and 50 μL membranes. Assay plates wereincubated for 60 minutes at 37° C. Binding reactions were terminated byrapid filtration over GF/B glass fiber filter plates (PerkinElmer Inc.,Wellesley, Mass.) pre-treated in 1% BSA. Filter plates were rinsed threetimes with wash buffer (10 mM HEPES) to remove unbound radioactivity.The plates were then air dried and 50 μL Microscint-20 liquidscintillation fluid (PerkinElmer Inc., Wellesley, Mass.) was added toeach well. The plates were then counted in a PerkinElmer Topcount liquidscintillation counter (PerkinElmer Inc., Wellesley, Mass.). Binding datawere analyzed by nonlinear regression analysis with the GraphPad PrismSoftware package (GraphPad Software, Inc., San Diego, Calif.) using theone-site competition model. K_(i) values for test compounds werecalculated from observed IC₅₀ values and the K_(D) value of theradioligand using the Cheng-Prusoff equation (Cheng Y; Prusoff W H.(1973) Biochemical Pharmacology, 22(23):3099-108). K_(i) values wereconverted to pK_(i) values to determine the geometric mean and 95%confidence intervals. These summary statistics were then converted backto K_(i) values for data reporting.

[0749] In this assay, a lower K_(i) value indicates that the testcompound has a higher binding affinity for the receptor tested.Exemplified compound that were tested in this assay typically were foundto have a K_(i) value of less than about 300 nM for the M₃ muscarinicreceptor. For example, the compounds of Examples 3 and 6 were identifiedas having K_(i) values of less than 10 nM.

Assay Test Procedure C Whole-cell cAMP Flashplate Assay in CHO CellLines Heterologously Expressing Human β₁, β₂ or β₃ Adrenergic Receptors

[0750] cAMP assays were performed in a radioimmunoassay format using theFlashplate Adenylyl Cyclase Activation Assay System with [¹²⁵I]-cAMP(NEN SMP004, PerkinElmer Life Sciences Inc., Boston, Mass.), accordingto the manufacturers instructions. For the determination of β receptoragonist potency (EC₅₀), CHO-K1 cell lines stably expressing cloned humanβ₁, β₂ or β₃ adrenergic receptors were grown to near confluency in HAM'sF-12 media supplemented with 10% FBS and Geneticin (250 μg/mL). Cellswere rinsed with PBS and detached in dPBS (Dulbecco's Phosphate BufferedSaline, without CaCl₂ and MgCl₂) containing 2 mM EDTA or Trypsin-EDTAsolution (0.05% trypsin/0.53 mM EDTA). After counting cells in Coultercell counter, cells were pelleted by centrifugation at 1,000 rpm andre-suspended in stimulation buffer containing IBMX (PerkinElmer Kit)pre-warmed to room temperature to a concentration of 1.6×10⁶ to 2.8×10⁶cells/mL. About 60,000 to 80,000 cells per well were used in this assay.Test compounds (10 mM in DMSO) were diluted into PBS containing 0.1% BSAin Beckman Biomek-2000 and tested at 11 different concentrations rangingfrom 100 μM to 1 pM. Reactions were incubated for 10 min at 37° C. andstopped by adding 100 μL of cold detection buffer containing [¹²⁵I]-cAMP(NEN SMP004, PerkinElmer Life Sciences, Boston, Mass.). The amount ofcAMP produced (pmol/well) was calculated based on the counts observedfor the samples and cAMP standards as described in the manufacturer'suser manual. Data were analyzed by nonlinear regression analysis withthe GraphPad Prism Software package (GraphPad Software, Inc., San Diego,Calif.) with the sigmoidal equation. The Cheng-Prusoff equation (ChengY, and Prusoff W H., Biochemical Pharmacology, 1973, 22, 23, 3099-108)was used to calculate the EC50 values.

[0751] In this assay, a lower EC₅₀ value indicates that the testcompound has a higher functional activity at the receptor tested.Exemplified compound that were tested in this assay typically were foundto have a EC₅₀ value of less than about 300 nM for the β₂ adrenergicreceptor. For example, the compounds of Examples 3 and 6 were identifiedas having EC₅₀ values of less than 10 nM.

[0752] If desired, the receptor subtype selectivity for a test compoundcan be calculated as the ratio of EC₅₀(β₁)/EC₅₀(β₂) orEC₅₀(β₃)/EC₅₀(β₂). Typically, compounds of this invention demonstratedgreater functional activity at the β₂ adrenergic receptor compared tothe β₁ or β₃ adrenergic receptor, i.e. EC₅₀(β₁) or EC₅₀(β₃) is typicallygreater than EC₅₀(β₂). Generally, compounds having selectivity for theβ₂ adrenergic receptor over the β₁ or β₃ adrenergic receptors arepreferred; especially compounds having a selectivity greater than about5; and in particular, greater than about 10. By way of example, thecompounds of Examples 3 and 6 had ratios of EC₅₀(β₁)/EC₅₀(β₂) greaterthan 10.

Assay Test Procedure D Functional Assays of Antagonism for MuscarinicReceptor Subtypes

[0753] A. Blockade of Agonist-Mediated Inhibition of cAMP Accumulation

[0754] In this assay, the functional potency of a test compound isdetermined by measuring the ability of the test compound to blockoxotremorine-inhibition of forskolin-mediated cAMP accumulation inCHO-K1 cells expressing the hM₂ receptor. cAMP assays are performed in aradioimmunoassay format using the Flashplate Adenylyl Cyclase ActivationAssay System with ¹²⁵I-cAMP (NEN SMP004B, PerkinElmer Life SciencesInc., Boston, Mass.), according to the manufacturer's instructions.Cells are rinsed once with dPBS and lifted with Trypsin-EDTA solution(0.05% trypsin/0.53 mM EDTA) as described in the Cell Culture andMembrane Preparation section above. The detached cells are washed twiceby centrifugation at 650×g for five minutes in 50 mL dPBS. The cellpellet is then re-suspended in 10 mL dPBS, and the cells are countedwith a Coulter Z1 Dual Particle Counter (Beckman Coulter, Fullerton,Calif.). The cells are centrifuged again at 650×g for five minutes andre-suspended in stimulation buffer to an assay concentration of1.6×10⁶-2.8×10⁶ cells/mL.

[0755] The test compound is initially dissolved to a concentration of400 μM in dilution buffer (dPBS supplemented with 1 mg/mL BSA (0.1%)),and then serially diluted with dilution buffer to final molarconcentrations ranging from 100 μM to 0.1 nM. Oxotremorine is diluted ina similar manner.

[0756] To measure oxotremorine inhibition of adenylyl cyclase (AC)activity, 25 μL forskolin (25 μM final concentration diluted in dPBS),25 μL diluted oxotremorine, and 50 μL cells are added to agonist assaywells. To measure the ability of a test compound to blockoxotremorine-inhibited AC activity, 25 μL forskolin and oxotremorine (25μM and 5 μM final concentrations, respectively, diluted in dPBS), 25 μLdiluted test compound, and 50 μL cells are added to remaining assaywells.

[0757] Reactions are incubated for 10 minutes at 37° C. and stopped byaddition of 100 μL ice-cold detection buffer. Plates are sealed,incubated overnight at room temperature and counted the next morning ona PerkinElmer TopCount liquid scintillation counter (PerkinElmer Inc.,Wellesley, Mass.). The amount of cAMP produced (pmol/well) is calculatedbased on the counts observed for the samples and cAMP standards, asdescribed in the manufacturer's user manual. Data is analyzed bynonlinear regression analysis with the GraphPad Prism Software package(GraphPad Software, Inc., San Diego, Calif.) using the non-linearregression, one-site competition equation. The Cheng-Prusoff equation isused to calculate the K_(i), using the EC₅₀ of the oxotremorineconcentration-response curve and the oxotremorine assay concentration asthe K_(D) and [L], respectively.

[0758] In this assay, a lower K_(i) value indicates that the testcompound has a higher functional activity at the receptor tested.Exemplified compound that were tested in this assay typically were foundto have a K_(i) value of less than about 300 nM for blockade ofoxotremorine-inhibition of forskolin-mediated cAMP accumulation inCHO-K1 cells expressing the hM₂ receptor. For example, the compound ofExample 3 was identified as having a K_(i) value of less than 10 nM.

[0759] B. Blockade of Agonist-Mediated [³⁵S]GTPγS Binding

[0760] In a second functional assay, the functional potency of testcompounds can be determined by measuring the ability of the compounds toblock oxotremorine-stimulated [³⁵S]GTPγS_binding in CHO-K1 cellsexpressing the hM₂ receptor.

[0761] At the time of use, frozen membranes were thawed and then dilutedin assay buffer with a final target tissue concentration of 5-10 μgprotein per well. The membranes were briefly homogenized using aPolytron PT-2100 tissue disrupter and then added to the assay plates.

[0762] The EC₉₀ value (effective concentration for 90% maximal response)for stimulation of [³⁵S]GTPγS binding by the agonist oxotremorine wasdetermined in each experiment.

[0763] To determine the ability of a test compound to inhibitoxotremorine-stimulated [³⁵S]GTPγS binding, the following was added toeach well of 96 well plates: 25 μL of assay buffer with [³⁵S]GTPγS (0.4nM), 25 μL of oxotremorine(EC₉₀) and GDP (3 uM), 25 μL of diluted testcompound and 25 μL CHO cell membranes expressing the hM₂ receptor. Theassay plates were then incubated at 37° C. for 60 minutes. The assayplates were filtered over 1% BSA-pretreated GF/B filters using aPerkinElmer 96-well harvester. The plates were rinsed with ice-cold washbuffer for 3×3 seconds and then air or vacuum dried. Microscint-20scintillation liquid (50 μL) was added to each well, and each plate wassealed and radioactivity counted on a Topcounter (PerkinElmer). Datawere analyzed by nonlinear regression analysis with the GraphPad PrismSoftware package (GraphPad Software, Inc., San Diego, Calif.) using thenon-linear regression, one-site competition equation. The Cheng-Prusoffequation was used to calculate the K_(i), using the IC₅₀ values of theconcentration-response curve for the test compound and the oxotremorineconcentration in the assay as the K_(D) and [L], ligand concentration,respectively.

[0764] In this assay, a lower K_(i) value indicates that the testcompound has a higher functional activity at the receptor tested.Exemplified compound that were tested in this assay typically were foundto have a K_(i) value of less than about 300 nM for blockade ofoxotremorine-stimulated [³⁵S]GTPγS_binding in CHO-K1 cells expressingthe hM₂ receptor. For example, the compound of Example 3 was identifiedas having a K_(i) value of less than 10 nM.

[0765] C. Blockade of Agonist-Mediated Calcium Release via FLIPR Assays

[0766] Muscarinic receptor subtypes (M₁, M₃ and M₅ receptors), whichcouple to G_(q) proteins, activate the phospholipase C (PLC) pathwayupon agonist binding to the receptor. As a result, activated PLChydrolyzes phosphatyl inositol diphosphate (PIP₂) to diacylglycerol(DAG) and phosphatidyl-1,4,5-triphosphate (IP₃), which in turn generatescalcium release from intracellular stores, i.e., endoplasmic andsarcoplasmic reticulum. The FLIPR (Molecular Devices, Sunnyvale, Calif.)assay capitalizes on this increase in intracellular calcium by using acalcium sensitive dye (Fluo-4AM, Molecular Probes, Eugene, Oreg.) thatfluoresces when free calcium binds. This fluorescence event is measuredin real time by the FLIPR, which detects the change in fluorescence froma monolayer of cells cloned with human M₁ and M₃, and chimpanzee M₅receptors. Antagonist potency can be determined by the ability ofantagonists to inhibit agonist-mediated increases in intracellularcalcium.

[0767] For FLIPR calcium stimulation assays, CHO cells stably expressingthe hM₁, hM₃ and cM₅ receptors are seeded into 96-well FLIPR plates thenight before the assay is done. Seeded cells are washed twice byCellwash (MTX Labsystems, Inc.) with FLIPR buffer (10 mM HEPES, pH 7.4,2 mM calcium chloride, 2.5 mM probenecid in Hank's Buffered SaltSolution (HBSS) without calcium and magnesium) to remove growth mediaand leaving 50 μL/well of FLIPR buffer. The cells are then incubatedwith 50 μL/well of 4 μM FLUO-4AM (a 2× solution was made) for 40 minutesat 37° C., 5% carbon dioxide. Following the dye incubation period, cellsare washed two times with FLIPR buffer, leaving a final volume of 50μL/well.

[0768] To determine antagonist potency, the dose-dependent stimulationof intracellular Ca²⁺ release for oxotremorine is first determined sothat antagonist potency can later be measured against oxotremorinestimulation at an EC₉₀ concentration. Cells are first incubated withcompound dilution buffer for 20 minutes, followed by agonist addition,which is performed by the FLIPR. An EC₉₀ value for oxotremorine isgenerated according to the method detailed in the FLIPR measurement anddata reduction section below, in conjunction with the formulaEC_(F)=((F/100-F){circumflex over ( )}1/H)*EC₅₀. An oxotremorineconcentration of 3×EC_(F) is prepared in stimulation plates such that anEC₉₀ concentration of oxotremorine is added to each well in theantagonist inhibition assay plates.

[0769] The parameters used for the FLIPR are: exposure length of 0.4seconds, laser strength of 0.5 watts, excitation wavelength of 488 nm,and emission wavelength of 550 nm. Baseline is determined by measuringthe change in fluorescence for 10 seconds prior to addition of agonist.Following agonist stimulation, the FLIPR continuously measured thechange of fluorescence every 0.5 to 1 second for 1.5 minutes to capturethe maximum fluorescence change.

[0770] The change of fluorescence is expressed as maximum fluorescenceminus baseline fluorescence for each well. The raw data is analyzedagainst the logarithm of drug concentration by nonlinear regression withGraphPad Prism (GraphPad Software, Inc., San Diego, Calif.) using thebuilt-in model for sigmoidal dose-response. Antagonist K_(i) values aredetermined by Prism using the oxotremorine EC₅₀ value as the K_(D) andthe oxotremorine EC₉₀ for the ligand concentration according to theCheng-Prusoff equation (Cheng & Prusoff, 1973).

[0771] In this assay, a lower K_(i) value indicates that the testcompound has a higher functional activity at the receptor tested.Exemplified compound that were tested in this assay typically were foundto have a K_(i) value of less than about 300 nM for blockade ofagonist-mediated calcium release in CHO cells stably expressing the hM₁,hM₃ and CM₅ receptors. For example, the compound of Example 3 wasidentified as having a K_(i) value of less than 10 nM for the hM₁, hM₃and cM₅ receptors.

Assay Test Procedure E Whole-cell cAMP Flashplate Assay With a LungEpithelial Cell Line Endogenously Expressing Human β₂ AdrenergicReceptor

[0772] For the determination of agonist potencies and efficacies(intrinsic activities) in a cell line expressing endogenous levels ofthe β₂ adrenergic receptor, a human lung epithelial cell line (BEAS-2B)was used (ATCC CRL-9609, American Type Culture Collection, Manassas,Va.) (January B, et al., British Journal of Pharmacology, 1998, 123, 4,701-11). Cells were grown to 75-90% confluency in complete, serum-freemedium (LHC-9 MEDIUM containing Epinephrine and Retinoic Acid, cat #181-500, Biosource International, Camarillo, Calif.). The day before theassay, medium was switched to LHC-8 (no epinephrine or retinoic acid,cat # 141-500, Biosource International, Camarillo, Calif.). cAMP assayswere performed in a radioimmunoassay format using the FlashplateAdenylyl Cyclase Activation Assay System with [¹²⁵I]-cAMP (NEN SMP004,PerkinElmer Life Sciences Inc., Boston, Mass.), according to themanufacturers instructions. On the day of the assay, cells were rinsedwith PBS, lifted by scraping with 5 mM EDTA in PBS, and counted. Cellswere pelleted by centrifugation at 1,000 rpm and re-suspended instimulation buffer pre-warmed to 37° C. at a final concentration of600,000 cells/mL. Cells were used at a final concentration of 100,000 to120,000 cells/well in this assay. Test compounds were serially dilutedinto assay buffer (75 mM Tris/HCl pH 7.4 at 25° C., 12.5 mM MgCl₂, 1 mMEDTA, 0.2% BSA) in Beckman Biomek-2000. Test compounds were tested inthe assay at 11 different concentrations, ranging from 10 μM to 10 pM.Reactions were incubated for 10 min at 37° C. and stopped by addition of100 μL of ice-cold detection buffer. Plates were sealed, incubated overnight at 4° C. and counted the next morning in a Topcount scintillationcounter (Packard BioScience Co., Meriden, Conn.). The amount of cAMPproduced per mL of reaction was calculated based on the counts observedfor samples and cAMP standards, as described in the manufacturer's usermanual. Data were analyzed by nonlinear regression analysis with theGraphPad Prism Software package (GraphPad Software, Inc., San Diego,Calif.) using the 4-parameter model for sigmoidal dose-response.

[0773] In this assay, a lower EC₅₀ value indicates that the testcompound has a higher functional activity at the receptor tested.Exemplified compound that were tested in this assay typically were foundto have a EC₅₀ value of less than about 300 nM for the β₂ adrenergicreceptor. For example, the compounds of Examples 3 and 6 were identifiedas having EC₅₀ values of less than 10 nM.

[0774] If desired, test compound efficacy (% Eff) was calculated fromthe ratio of the observed Emax (TOP of the fitted curve) and the maximalresponse obtained for isoproterenol dose response curve and wasexpressed as % Eff relative to isoproterenol. Exemplified compoundstested in this assay typically demonstrated a % Eff greater than about40.

Assay Test Procedure F Duration of Bronchoprotection in Guinea PigModels of Acetylcholine-Induced or Histamine-Induced Bronchoconstriction

[0775] These in vivo assays were used to assess the bronchoprotectiveeffects of test compounds exhibiting both muscarinic receptor antagonistand β₂ adrenergic receptor agonist activity. To isolate muscarinicantagonist activity in the acetylcholine-induced bronchoconstrictionmodel, the animals were administered propanolol, a compound that blocksβ receptor activity, prior to the administration of acetylcholine.Duration of bronchoprotection in the histamine-inducedbronchoconstriction model reflects β₂ adrenergic receptor agonistactivity.

[0776] Groups of 6 male guinea pigs (Duncan-Hartley (HsdPoc:DH) Harlan,Madison, Wis.) weighing between 250 and 350 g were individuallyidentified by cage cards. Throughout the study, animals were allowedaccess to food and water ad libitum.

[0777] Test compounds were administered via inhalation over 10 minutesin a whole-body exposure dosing chamber (R&S Molds, San Carlos, Calif.).The dosing chambers were arranged so that an aerosol was simultaneouslydelivered to 6 individual chambers from a central manifold. Guinea pigswere exposed to an aerosol of a test compound or vehicle (WFI). Theseaerosols were generated from aqueous solutions using an LC StarNebulizer Set (Model 22F51, PARI Respiratory Equipment, Inc. Midlothian,Va.) driven by a mixture of gases (CO₂=5%, O₂=21% and N₂=74%) at apressure of 22 psi. The gas flow through the nebulizer at this operatingpressure was approximately 3 L/minute. The generated aerosols weredriven into the chambers by positive pressure. No dilution air was usedduring the delivery of aerosolized solutions. During the 10 minutenebulization, approximately 1.8 mL of solution was nebulized. This valuewas measured gravimetrically by comparing pre-and post-nebulizationweights of the filled nebulizer.

[0778] The bronchoprotective effects of test compounds administered viainhalation were evaluated using whole body plethysmography at 1.5, 24,48 and 72 hours post-dose.

[0779] Forty-five minutes prior to the start of the pulmonaryevaluation, each guinea pig was anesthetized with an intramuscularinjection of ketamine (43.75 mg/kg), xylazine (3.50 mg/kg) andacepromazine (1.05 mg/kg). After the surgical site was shaved andcleaned with 70% alcohol, a 2-3 cm midline incision of the ventralaspect of the neck was made. Then, the jugular vein was isolated andcannulated with a saline-filled polyethylene catheter (PE-50, BectonDickinson, Sparks, Md.) to allow for intravenous infusions ofacetylcholine (Ach) or histamine in saline. The trachea was thendissected free and cannulated with a 14G teflon tube (#NE-014, SmallParts, Miami Lakes, Fla.). If required, anesthesia was maintained byadditional intramuscular injections of the aforementioned anestheticmixture. The depth of anesthesia was monitored and adjusted if theanimal responds to pinching of its paw or if the respiration rate wasgreater than 100 breaths/minute.

[0780] Once the cannulations were completed, the animal was placed intoa plethysmograph (#PLY3114, Buxco Electronics, Inc., Sharon, Conn.) andan esophageal pressure cannula (PE-160, Becton Dickinson, Sparks, Md.)was inserted to measure pulmonary driving pressure (pressure). Theteflon tracheal tube was attached to the opening of the plethysmographto allow the guinea pig to breathe room air from outside the chamber.The chamber was then sealed. A heating lamp was used to maintain bodytemperature and the guinea pig's lungs were inflated 3 times with 4 mLof air using a 10 mL calibration syringe (#5520 Series, Hans Rudolph,Kansas City, Mo.) to ensure that the lower airways did not collapse andthat the animal did not suffer from hyperventilation.

[0781] Once it was determined that baseline values were within the rangeof 0.3-0.9 mL/cm H₂O for compliance and within the range of 0.1-0.199 cmH₂O/mL per second for resistance, the pulmonary evaluation wasinitiated. A Buxco pulmonary measurement computer program enabled thecollection and derivation of pulmonary values.

[0782] Starting this program initiated the experimental protocol anddata collection. The changes in volume over time that occur within theplethysmograph with each breath were measured via a Buxco pressuretransducer. By integrating this signal over time, a measurement of flowwas calculated for each breath. This signal, together with the pulmonarydriving pressure changes, which were collected using a Sensym pressuretransducer (#TRD4100), was connected via a Buxco (MAX 2270) preamplifierto a data collection interface (#'s SFT3400 and SFT3813). All otherpulmonary parameters were derived from these two inputs.

[0783] Baseline values were collected for 5 minutes, after which timethe guinea pigs were challenged with Ach or histamine. When evaluatingthe muscarinic antagonist effects, propanolol (5 mg/Kg, iv)(Sigma-Aldrich, St. Louis, Mo.) was administered 15 minutes prior tochallenge with Ach. Ach (Sigma-Aldrich, St. Louis, Mo.) (0.1 mg/mL) wasinfused intravenously for 1 minute from a syringe pump (sp210iw, WorldPrecision Instruments, Inc., Sarasota, Fla.) at the following doses andprescribed times from the start of the experiment: 1.9 μg/minute at 5minutes, 3.8 μg/minute at 10 minutes, 7.5 μg/minute at 15 minutes, 15.0μg/minute at 20 minutes, 30 μg/minute at 25 minutes and 60 μg/minute at30 minutes. Alternatively, bronchoprotection of test compounds wasassessed in the acetylcholine challenge model without pretreatment witha beta blocking compound.

[0784] When evaluating the β₂ adrenergic receptor agonist effects oftest compounds, histamine (25 μg/mL) (Sigma-Aldrich, St. Louis, Mo.) wasinfused intravenously for 1 minute from a syringe pump at the followingdoses and prescribed times from the start of the experiment: 0.5μg/minute at 5 minutes, 0.9 μg/minute at 10 minutes, 1.9 μg/minute at 15minutes, 3.8 μg/minute at 20 minutes, 7.5 μg/minute at 25 minutes and 15μg/minute at 30 minutes. If resistance or compliance had not returned tobaseline values at 3 minutes following each Ach or histamine dose, theguinea pig's lungs were inflated 3 times with 4 mL of air from a 10 mLcalibration syringe. Recorded pulmonary parameters include respirationfrequency (breaths/minute), compliance (mL/cm H₂O) and pulmonaryresistance (cm H₂O/mL per second). Once the pulmonary functionmeasurements were completed at minute 35 of this protocol, the guineapig was removed from the plethysmograph and euthanized by carbon dioxideasphyxiation.

[0785] The data were evaluated in one of two ways:

[0786] (a) Pulmonary resistance (R_(L), cm H₂O/mL per second) wascalculated from the ratio of “change in pressure” to “the change inflow.” The R_(L) response to ACh (60 μg/min, IH) was computed for thevehicle and the test compound groups. The mean ACh response invehicle-treated animals, at each pre-treatment time, was calculated andused to compute % inhibition of ACh response, at the correspondingpre-treatment time, at each test compound dose. Inhibition dose-responsecurves for ‘R_(L)’ were fitted with a four parameter logistic equationusing GraphPad Prism, version 3.00 for Windows (GraphPad Software, SanDiego, Calif.) to estimate bronchoprotective ID₅₀ (dose required toinhibit the ACh (60 μg/min) bronchocontrictor response by 50%). Theequation used was as follows:

Y=Min+(Max−Min)/(1+10^(((log ID50-X)*Hillslope)))

[0787] where X is the logarithm of dose, Y is the response (% Inhibitionof ACh induced increase in R_(L)). Y starts at Min and approachesasymptotically to Max with a sigmoidal shape.

[0788] (b) The quantity PD₂, which is defined as the amount of Ach orhistamine needed to cause a doubling of the baseline pulmonaryresistance, was calculated using the pulmonary resistance values derivedfrom the flow and the pressure over a range of Ach or histaminechallenges using the following equation (derived from the equation usedto calculate PC₂₀ values in the clinic (see Am. Thoracic Soc, 2000):${PD}_{2} = {{antilog}\left\lbrack {{\log \quad C_{1}} + \frac{\left( {{\log \quad C_{2}} - {\log \quad C_{1}}} \right)\left( {{2R_{0}} - R_{1}} \right)}{R_{2} - R_{1}}} \right\rbrack}$

[0789] where:

[0790] C₁=concentration of Ach or histamine preceding C₂

[0791] C₂=concentration of Ach or histamine resulting in at least a2-fold increase in pulmonary resistance (R_(L))

[0792] R₀=Baseline R_(L) value

[0793] R₁=R_(L) value after C₁

[0794] R₂=R_(L) value after C₂

[0795] Statistical analysis of the data was performed using a twotailed—Students t-test. A P-value <0.05 was considered significant.

[0796] Exemplified compounds that were tested in this assay typicallyproduced a dose-dependent bronchoprotective effect against MCh-inducedbronchoconstriction and His-induced bronchoconstriction. Generally, testcompounds having a potency (ID₅₀ at 1.5 h post-dose) of less than about300 μg/mL for ACh-induced bronchoconstriction and less than about 300μg/mL for His-induced bronchoconstriction in this assay are preferred.For example, the compounds of Examples 3 and 6 were found to have anID₅₀ less than about 100 μg/mL for ACh-induced bronchoconstriction andan ID₅₀ less than about 100 μg/mL for His-induced bronchoconstriction at1.5 hours post-dose.

[0797] Additionally, test compounds having a duration (PD T_(1/2)) ofbrochoprotective activity of at least about 24 hours in this assay aregenerally preferred. By way of example, the compounds of Examples 3 and6 were identified as having a PD T_(1/2) of at least about 24 hourspost-dose.

Assay Test Procedure G Einthoven Model for Measuring Changes inVentilation in Guinea Pigs

[0798] The bronchodilator activity of test compounds was evaluated in ananesthetized guinea pig model (the Einthoven model), which usesventilation pressure as a surrogate measure of airway resistance. See,for example, Einthoven (1892) Pfugers Arch. 51: 367-445; and Mohammed etal. (2000) Pulm Pharmacol Ther.13(6):287-92. In this model, muscarinicantagonist and β₂ agonist activity was assessed by determining theprotective effects against methacholine (MCh) and histamine(His)-induced bronchoconstriction.

[0799] This assay was conducted using Duncan-Hartley guinea pigs(Harlan, Indianapolis, Ind.), weighing between 300 and 400 g.

[0800] The test compound or vehicle (i.e., sterile water) was dosed byinhalation (1H) over a 10 minute time period in a whole body exposuredosing chamber (R+S Molds, San Carlos, Calif.) using 5 mL of dosingsolution. Animals were exposed to an aerosol, which was generated froman LC Star Nebulizer Set (Model 22F51, PARI Respiratory Equipment, Inc.Midlothian, Va.) driven by Bioblend a mixture of gasses (5% CO₂; 21% O₂;and 74% N₂) at a pressure of 22 psi. Pulmonary function was evaluated atvarious time-points after inhalation dosing.

[0801] Forty five minutes prior to the start of pulmonary functionevaluation, the guinea pigs were anesthetized with an intramuscular (IM)injection of a mixture of ketamine (13.7 mg/kg/xylazine (3.5mg/kg)/acepromazine (1.05 mg/kg). A supplemental dose of this mixture(50% of initial dose) was administered as needed. The jugular vein andcarotid artery were isolated and cannulated with saline-filledpolyethylene catheters (micro-renathane and PE-50, respectively, BecktonDickinson, Sparks, Md.). The carotid artery was connected to a pressuretransducer to allow the measurement of blood pressure and the jugularvein cannula was used for IV injection of either MCh or His. The tracheawas then dissected free and cannulated with a 14G needle (#NE-014, SmallParts, Miami Lakes, Fla.). Once the cannulations were complete, theguinea pigs were ventilated using a respirator (Model 683, HarvardApparatus, Inc., Mass.) set at a stroke volume of 1 mL/100 g body weightbut not exceeding 2.5 mL volume, and at a rate of 100 strokes perminute. Ventilation pressure (VP) was measured in the tracheal cannulausing a Biopac transducer that was connected to a Biopac (TSD 137C)pre-amplifier. Body temperature was maintained at 37° C. using a heatingpad. Prior to initiating data collection, pentobarbital (25 mg/kg) wasadministered intraperitoneally (IP) to suppress spontaneous breathingand obtain a stable baseline. The changes in VP were recorded on aBiopac Windows data collection interface. Baseline values were collectedfor at least 5 minutes, after which time guinea pigs were challenged IVnon-cumulatively with 2-fold incremental doses of the bronchoconstrictor(MCh or His). When MCh was used as the bronchoconstrictor agent, animalswere pre-treated with propranolol (5 mg/kg, IV) to isolate theantimuscarinic effects of the test compound. Changes in VP were recordedusing the Acknowledge Data Collection Software (Santa Barbara, Calif.).After the completion of study, the animals were euthanized.

[0802] Change in VP was measured in cm of water. Change in VP (cmH₂O)=peak pressure (after bronchoconstrictor challenge)−peak baselinepressure. The dose-response curve to MCh or His was fitted to a fourparameter logistic equation using GraphPad Prism, version 3.00 forWindows (GraphPad Software, San Diego, Calif.) The equation used was asfollows:

Y=Min+(Max−Min)/(1+10^(((log ID50-X)*Hillislope)))

[0803] where X is the logarithm of dose, Y is the response. Y starts atMin and approaches asymptotically to Max with a sigmoidal shape.

[0804] The percent inhibition of the bronchoconstrictor response to asubmaximal dose of MCh or His was calculated at each dose of the testcompound using the following equation: % Inhibition ofresponse=100−((peak pressure (after bronchoconstrictor challenge,treated)−peak baseline pressure (treated)*100% / (peak pressure (afterbronchoconstrictor challenge, water)−peak baseline pressure (water)).Inhibition curves were fitted using the four parameter logistic equationfrom GraphPad software. ID₅₀ (dose required to produce 50% inhibition ofthe bronchoconstrictor response) and Emax (maximal inhibition) were alsoestimated wherever appropriate.

[0805] The magnitude of bronchoprotection at different time-points afterinhalation of the test compound was used to estimate the pharmacodynamichalf-life (PD T_(1/2)). PD T_(1/2) was determined using a non-linearregression fit using a one-phase exponential decay equation (GraphPadPrism, Version 4.00): Y=Span*exp(−K*X)+Plateau; Starts at Span+Plateauand decays to Plateau with a rate constant K. The PD T_(1/2)=0.69/K.Plateau was constrained to 0.

[0806] Exemplified compounds that were tested in this assay typicallyproduced a dose-dependent bronchoprotective effect against MCh-inducedbronchoconstriction and His-induced bronchoconstriction. Generally, testcompounds having an ID₅₀ less than about 300 μg/mL for MCh-inducedbronchoconstriction and an ID₅₀ less than about 300 μg/mL forHis-induced bronchoconstriction at 1.5 hours post-dose in this assay arepreferred. Additionally, test compounds having a duration (PD T_(1/2))of brochoprotective activity of at least about 24 hours in this assayare generally preferred.

Assay Test Procedure H Inhalation Guinea Pig Salivation Assay

[0807] Guinea pigs (Charles River, Wilmington, Mass.) weighing 200-350 gwere acclimated to the in-house guinea pig colony for at least 3 daysfollowing arrival. Test compound or vehicle were dosed via inhalation(IH) over a 10 minute time period in a pie shaped dosing chamber (R+SMolds, San Carlos, Calif.). Test solutions were dissolved in sterilewater and delivered using a nebulizer filled with 5.0 mL of dosingsolution. Guinea pigs were restrained in the inhalation chamber for 30minutes. During this time, guinea pigs were restricted to an area ofapproximately 110 sq. cm. This space was adequate for the animals toturn freely, reposition themselves, and allow for grooming. Following 20minutes of acclimation, guinea pigs were exposed to an aerosol generatedfrom a LS Star Nebulizer Set (Model 22F51, PARI Respiratory Equipment,Inc. Midlothian, Va.) driven by house air at a pressure of 22 psi. Uponcompletion of nebulization, guinea pigs were evaluated at 1.5, 6, 12,24, 48, or 72 hrs after treatment.

[0808] Guinea pigs were anesthetized one hour before testing with anintramuscular (IM) injection of a mixture of ketamine 43.75 mg/kg,xylazine 3.5 mg/kg, and acepromazine 1.05 mg/kg at an 0.88 mL/kg volume.Animals were placed ventral side up on a heated (37° C.) blanket at a 20degree incline with their head in a downward slope. A 4-ply 2×2 inchgauze pad (Nu-Gauze General-use sponges, Johnson and Johnson, Arlington,Tex.) was inserted in the guinea pig's mouth. Five minutes later, themuscarinic agonist pilocarpine (3.0 mg/kg, s.c.) was administered andthe gauze pad was immediately discarded and replaced by a newpre-weighed gauze pad. Saliva was collected for 10 minutes, at whichpoint the gauze pad was weighed and the difference in weight recorded todetermine the amount of accumulated saliva (in mg). The mean amount ofsaliva collected for animals receiving the vehicle and each dose of testcompound was calculated. The vehicle group mean was considered to be100% salivation. Results were calculated using result means (n=3 orgreater). Confidence intervals (95%) were calculated for each dose ateach time point using two-way ANOVA. This model is a modified version ofthe procedure described in Rechter, “Estimation of anticholinergic drugeffects in mice by antagonism against pilocarpine-induced salivation”Ata Pharmacol Toxicol, 1996, 24:243-254.

[0809] The mean weight of saliva in vehicle-treated animals, at eachpre-treatment time, was calculated and used to compute % inhibition ofsalivation, at the corresponding pre-treatment time, at each dose. Theinhibition dose-response data were fitted to a a four parameter logisticequation using GraphPad Prism, version 3.00 for Windows (GraphPadSoftware, San Diego, Calif.) to estimate anti-sialagogue ID₅₀ (doserequired to inhibit 50% of pilocarpine-evoked salivation). The equationused was as follows:

Y=Min+(Max−Min)/(1+10^(((log ID50-X)*Hillislope)))

[0810] where X is the logarithm of dose, Y is the response (% inhibitionof salivation). Y starts at Min and approaches asymptotically to Maxwith a sigmoidal shape.

[0811] The ratio of the anti-sialagogue ID₅₀ to bronchoprotective ID₅₀was used to compute the apparent lung-selectivity index of the testcompound. Generally, compounds having an apparent lung-selectivity indexgreater than about 5 are preferred. In this assay, the compound ofExample 3 was identified as having an apparent lung-selectivity indexgreater than 5.

[0812] While the present invention has been described with reference tospecific aspects or embodiments thereof, it will be understood by thoseof ordinary skilled in the art that various changes can be made orequivalents can be substituted without departing from the true spiritand scope of the invention. Additionally, to the extent permitted byapplicable patent statues and regulations, all publications, patents andpatent applications cited herein are hereby incorporated by reference intheir entirety to the same extent as if each document had beenindividually incorporated by reference herein.

What is claimed is:
 1. A library comprising compounds of formula I:

wherein: a is 0 or an integer of from 1 to 3; each R¹ is independentlyselected from (1-4C)alkyl, (2-4C)alkenyl, (2-4C)alkynyl,(3-6C)cycloalkyl, cyano, halo, —OR^(1a), —C(O)OR^(1b), —SR^(1c),—S(O)R^(1d), —S(O)₂R^(1e) and —NR^(1f)R^(1g); each of R^(1a), R^(1b),R^(1c), R^(1d), R^(1e), R^(1f) and R^(1g) is independently hydrogen,(1-4C)alkyl or phenyl-(1-4C)alkyl; b is 0 or an integer of from 1 to 3;each R² is independently selected from (1-4C)alkyl, (2-4C)alkenyl,(2-4C)alkynyl, (3-6C)cycloalkyl, cyano, halo, —OR^(2a), —C(O)OR^(2b),—SR^(2c), —S(O)R^(2d), —S(O)₂R^(2e) and —NR^(2f)R^(2g); each of R^(2a),R^(2b), R^(2c), R^(2d), R^(2e), R^(2f) and R^(2g) is independentlyhydrogen, (1-4C)alkyl or phenyl-(1-4C)alkyl; W is attached to the 3- or4-position with respect to the nitrogen atom in the piperidine ring andrepresents O or NW^(a); W^(a) is hydrogen or (1-4C)alkyl; c is 0 or aninteger of from 1 to 4; each R³ is independently selected from(1-4C)alkyl, (2-4C)alkenyl, (2-4C)alkynyl, (3-6C)cycloalkyl, cyano,halo, —OR^(3a), —C(O)OR^(3b), —SR^(3c), —S(O)R^(3d), S(O)₂R^(3e) and—NR^(3f)R^(3g); or two R³ groups are joined to form (1-3C)alkylene,(2-3C)alkenylene or oxiran-2,3-diyl; each of R^(3a), R^(3b), R^(3c),R^(3d), R^(3e), R^(3f) and R^(3g) is independently hydrogen or(1-4C)alkyl; R⁴ represents a divalent hydrocarbon group containing from4 to 28 carbon atoms and optionally containing from 1 to 10 heteroatomsselected independently from halo, oxygen, nitrogen and sulfur; R⁵represents hydrogen or (1-4C)alkyl; R⁶ is —NR^(6a)CR^(6b)(O) or—CR^(6c)R^(6d)OR^(6e) and R⁷ is hydrogen; or R⁶ and R⁷ together form—NR^(7a)C(O)—CR^(7b)═CR^(7c)—, —CR^(7d)═CR^(7e—C(O)—NR) ^(7f)—,—NR^(7g)C(O)—CR^(7h)R^(7i)—CR^(7j)R^(7k)— or—CR^(7l)R^(7m)—CR^(7n)R^(7o)—C(O)—NR^(7p); each of R^(6a), R^(6b),R^(6c), R^(6d) and R^(6e) is independently hydrogen or (1-4C)alkyl; andeach of R^(7a), R^(7b), R^(7c), R^(7d), R^(7e), R^(7f), R^(7g), R^(7h),R^(7i), R^(7j), R^(7k), R^(7l), R^(7m), R^(7n), R^(7o), and R^(7p) isindependently hydrogen or (1-4C)alkyl; or a salt or stereoisomerthereof.
 2. The library of claim 1, wherein the library contains acompound of formula I in which W is attached to the piperidine ring atthe 4-position with respect to the nitrogen atom.
 3. The library ofclaim 1, wherein the library contains a compound of formula I in whicha, b, and c are each 0, and W^(a) and R⁵ are both hydrogen.
 4. Thelibrary of claim 1, wherein the library contains a compound of formula Iin which W represents O.
 5. The library of claim 1, wherein the librarycontains a compound of formula I in which R⁶ is —NHCHO or —CH₂OH and R⁷is hydrogen; or R⁶ and R⁷ together form —NHC(O)—CH═CH—, —CH═CH—C(O)—NH—,—CH₂—CH₂—C(O)NH— or —NHC(O)—CH₂—CH₂—.
 6. The library of claim 1, whereinthe library contains a compound of formula I in which R⁴ is a divalentgroup of the formula:—(R^(4a))_(d)—(A¹)_(e)—(R^(4b))_(f)—Q—(R^(4c))_(g)—(A²)_(h)—(R^(4d))_(i)—wherein d, e, f, g, h and i are each independently selected from 0 and1; R^(4a), R^(4b), R^(4c) and R^(4d) are each independently selectedfrom (1-10C)alkylene, (2-10C)alkenylene and (2-10C)alkynylene, whereineach alkylene, alkenylene or alkynylene group is unsubstituted orsubstituted with from 1 to 5 substituents independently selected from(1-4C)alkyl, fluoro, hydroxy, phenyl and phenyl-(1-4C)alkyl; A¹ and A²are each independently selected from (3-7C)cycloalkylene,(6-10C)arylene, —O—(6-10C)arylene, (6-10C)arylene-O—,(2-9C)heteroarylene, —O—(2-9C) heteroarylene, (2-9C)heteroarylene-O— and(3-6C)heterocyclene, wherein each cycloalkylene is unsubstituted orsubstituted with from 1 to 4 substitutents selected independently from(1-4C)alkyl, and each arylene, heteroarylene or heterocyclene group isunsubstituted or substituted with from 1 to 4 substituents independentlyselected from halo, (1-4C)alkyl, (1-4C)alkoxy, —S-(1-4C)alkyl,—S(O)-(1-4C)alkyl, —S(O)₂-(1-4C)alkyl, —C(O)O(1-4C)alkyl, carboxy,cyano, hydroxy, nitro, trifluoromethyl and trifluoromethoxy; Q isselected from a bond, —O—, —C(O)O—, —OC(O)—, —S—, —S(O)—, —S(O)₂—,—N(Q^(a))C(O), —C(O)N(Q^(b))—, —N(Q^(c))S(O)₂—, —S(O)₂N(Q^(d))—,—N(Q^(e))C(O)N(Q^(f))—, —N(Q^(g))S(O)₂N(Q^(h))—, —OC(O)N(Q^(i))—,—N(Q^(j))C(O)O— and —N(Q^(k)); Q^(a), Q^(b), Q^(c), Q^(d), Q^(e), Q^(f),Q^(g), Q^(h), Q^(i), Q^(j) and Q^(k) are each independently selectedfrom hydrogen, (1-6C)alkyl, A³ and (1-4C)alkylene-A⁴, wherein the alkylgroup is unsubstituted or substituted with from 1 to 3 substituentsindependently selected from fluoro, hydroxy and (1-4C)alkoxy; ortogether with the nitrogen atom and the group R^(4b) or R^(4c) to whichthey are attached, form a 4-6 membered azacycloalkylene group; A³ and A⁴are each independently selected from (3-6C)cycloalkyl, (6-10C)aryl,(2-9C)heteroaryl and (3-6C)heterocyclyl, wherein each cycloalkyl isunsubstituted or substituted with from 1 to 4 substitutents selectedindependently from (1-4C)alkyl and each aryl, heteroaryl or heterocyclylgroup is unsubstituted or substituted with from 1 to 4 substituentsindependently selected from halo, (1-4C)alkyl and (1-4C)alkoxy.
 7. Thelibrary of claim 6, wherein the library contains a compound of formula Iin which R⁴ is a divalent group of the formula: —(R^(4a))_(d)— whereR^(4a) is (4-10C)alkylene.
 8. The library of claim 6, wherein thelibrary contains a compound of formula I in which R⁴ is —(CH₂)₈—,—(CH₂)₉, and —(CH₂)₁₀—.
 9. The library of claim 6, wherein the librarycontains a compound of formula I in which R⁴ is a divalent group of theformula: —(R^(4a))_(d)—(A²)_(h)—(R^(4d))_(i)—wherein R^(4a) is(1-10C)alkylene; A² is (6-10C)arylene or (2-9C)heteroarylene; and R^(4d)is (1-10C)alkylene.
 10. The library of claim 6, wherein the librarycontains a compound of formula I in which R⁴ is a divalent group of theformula: —(R^(4a))_(d)—(A²)_(h)—(R^(4d))_(i)—wherein Q is —O— or—N(Q^(k))—; Q^(k) is hydrogen or (1-3C)alkyl; R^(4a) is (1-10C)alkylene;A² is (6-10C)arylene or (2-9C)heteroarylene; and R^(4d) is(1-10C)alkylene.
 11. The library of claim 6, wherein the librarycontains a compound of formula I in which Q is —N(Q^(a))C(O)— or—C(O)N(Q^(b))—.
 12. The library of claim 1, wherein the library containsa compound of formula I in which R⁴ is selected from:

wherein m is an integer from 2 to 10; and n is an integer from 2 to 10;provided that m+n is an integer from 4 to 12;

wherein o is an integer from 2 to 7; and p is an integer from 1 to 6;provided that o+p is an integer from 3 to 8; and wherein thephen-1,4-ylene group is optionally substituted with from 1 to 4substituents independently selected from halo, (1-4C)alkyl,(1-4C)alkoxy, —S-(1-4C)alkyl, —S(O)-(1-4C)alkyl, —S(O)₂-(1-4C)alkyl,carboxy, cyano, hydroxy, nitro, trifluoromethyl and trifluoromethoxy;

wherein q is an integer from 2 to 6; r is an integer from 1 to 5; and sis an integer from 1 to 5; provided that q+r+s is an integer from 4 to8; and wherein the phen-1,4-ylene group is optionally substituted withfrom 1 to 4 substituents independently selected from halo, (1-4C)alkyl,(1-4C)alkoxy, —S-(1-4C)alkyl, —S(O)-(1-4C)alkyl, —S(O)₂-(1-4C)alkyl,—C(O)O(1-4C)alkyl, carboxy, cyano, hydroxy, nitro, trifluoromethyl andtrifluoromethoxy;

wherein t is an integer from 2 to 10; and u is an integer from 2 to 10;provided that t+u is an integer from 4 to 12;

wherein v is an integer from 2 to 7; and w is an integer from 1 to 6;provided that v+w is an integer from 3 to 8; and wherein thephen-1,4-ylene group is optionally substituted with from 1 to 4substituents independently selected from halo, (1-4C)alkyl,(1-4C)alkoxy, —S-(1-4C)alkyl, —S(O)-(1-4C)alkyl, —S(O)₂-(1-4C)alkyl,—C(O)O(1-4C)alkyl, carboxy, cyano, hydroxy, nitro, trifluoromethyl andtrifluoromethoxy; and

wherein x is an integer from 2 to 6; y is an integer from 1 to 5; and zis an integer from 1 to 5; provided that x+y+z is an integer from 4 to8; and wherein the phen-1,4-ylene group is optionally substituted withfrom 1 to 4 substituents independently selected from halo, (1-4C)alkyl,(1-4C)alkoxy, —S-(1-4C)alkyl, —S(O)-(1-4C)alkyl, —S(O)₂-(1-4C)alkyl,—C(O)O(1-4C)alkyl, carboxy, cyano, hydroxy, nitro, trifluoromethyl andtrifluoromethoxy.
 13. The library of claim 1, wherein the librarycontains a compound of formula II:

wherein W represents O or NW^(a); W^(a) is hydrogen or (1-4C)alkyl; R⁴represents a divalent hydrocarbon group containing from 4 to 28 carbonatoms and optionally containing from 1 to 10 heteroatoms selectedindependently from halo, oxygen, nitrogen and sulfur, or a salt orstereoisomer thereof.
 14. The library of claim 1, wherein the librarycontains a compound of formula III:

wherein W represents O or NW^(a); W^(a) is hydrogen or (1-4C)alkyl; R⁴represents a divalent hydrocarbon group containing from 4 to 28 carbonatoms and optionally containing from 1 to 10 heteroatoms selectedindependently from halo, oxygen, nitrogen and sulfur, or a salt orstereoisomer thereof.
 15. The library of claim 1, wherein the librarycontains a compound of formula IV:

wherein W represents O or NW^(a); W^(a) is hydrogen or (1-4C)alkyl; R⁴represents a divalent hydrocarbon group containing from 4 to 28 carbonatoms and optionally containing from 1 to 10 heteroatoms selectedindependently from halo, oxygen, nitrogen and sulfur, or a salt orstereoisomer thereof.
 16. A method of identifying a compound of formulaI having both β₂ adrenergic receptor agonist activity and muscarinicreceptor antagonist activity; the method comprising: evaluating thelibrary of any one of claims 1, 13, 14 or 15 to determine whether acompound of formula I in the library has both β₂ adrenergic receptoragonist activity and muscarinic receptor antagonist activity.
 17. Amethod of identifying a compound of formula I having both β₂ adrenergicreceptor agonist activity and muscarinic receptor antagonist activity;the method comprising: (a) preparing the library of any one of claims 1to 15; (b) evaluating the compounds of formula I in the library todetermine whether a compound has both β₂ adrenergic receptor agonistactivity and muscarinic receptor antagonist activity.
 18. A method ofidentifying a compound of formula I that binds to a β₂ adrenergicreceptor and a muscarinic receptor; the method comprising: evaluatingthe library of any one of claims 1, 13, 14 or 15 to determine whether acompound of formula I in the library has a K_(i) value of less thanabout 300 nM for a β₂ adrenergic receptor and a K_(i) value less than300 nM for a muscarinic receptor.
 19. A method of identifying a compoundof formula I that binds to a β₂ adrenergic receptor and a muscarinicreceptor; the method comprising: (a) preparing the library of any one ofclaims 1 to 15; and (b) evaluating the compounds of formula I in thelibrary to determine whether a compound has a K_(i) value of less thanabout 300 nM for a β₂ adrenergic receptor and a K_(i) value less than300 nM for a muscarinic receptor.
 20. A method of identifying a compoundof formula I that binds to a β₂ adrenergic receptor and a muscarinicreceptor; the method comprising: evaluating the library of any one ofclaims 1, 13, 14 or 15 to determine whether a compound of formula I inthe library has an EC₅₀ value of less than about 100 nM for a β₂adrenergic receptor and a K_(i) value less than 100 nM for a muscarinicreceptor.
 21. A method of identifying a compound of formula I that bindsto a β₂ adrenergic receptor and a muscarinic receptor; the methodcomprising: (a) preparing the library of any one of claims 1 to 15; (b)evaluating the compounds of formula I in the library to determinewhether a compound has a EC₅₀ value of less than about 100 nM for a β₂adrenergic receptor and a K_(i) value less than 100 nM for a muscarinicreceptor.
 22. A process for preparing a library comprising compounds offormula I:

wherein: a is 0 or an integer of from 1 to 3; each R¹ is independentlyselected from (1-4C)alkyl, (2-4C)alkenyl, (2-4C)alkynyl,(3-6C)cycloalkyl, cyano, halo, —OR^(1a), —C(O)OR^(1b), —SR^(1c),—S(O)R^(1d), —S(O)₂R^(1e) and —NR^(1f)R^(1g); each of R^(1a), R^(1b),R^(1c), R^(1d), R^(1e), R^(1f) and R^(1g) is independently hydrogen,(1-4C)alkyl or phenyl-(1-4C)alkyl; b is 0 or an integer of from 1 to 3;each R² is independently selected from (1-4C)alkyl, (2-4C)alkenyl,(2-4C)alkynyl, (3-6C)cycloalkyl, cyano, halo, —OR^(2a), —C(O)OR^(2b),—SR^(2c), —S(O)R^(2d), —S(O)₂R^(2e) and —NR^(2f)R^(2g); each of R^(2a),R^(2b), R^(2c), R^(2d), R^(2e), R^(2f) and R^(2g) is independentlyhydrogen, (1-4C)alkyl or phenyl-(1-4C)alkyl; W is attached to the 3- or4-position with respect to the nitrogen atom in the piperidine ring andrepresents O or NW^(a); W^(a) is hydrogen or (1-4C)alkyl; c is 0 or aninteger of from 1 to 4; each R³ is independently selected from(1-4C)alkyl, (2-4C)alkenyl, (2-4C)alkynyl, (3-6C)cycloalkyl, cyano,halo, —OR^(3a), —C(O)OR^(3b), —SR^(3c), —S(O)R^(3d), —S(O)₂R^(3e) and—NR^(3f)R^(3g); or two R³ groups are joined to form (1-3C)alkylene,(2-3C)alkenylene or oxiran-2,3-diyl; ach of R^(3a), R^(3b), R^(3c),R^(3d), R^(3e), R^(3f) and R^(3g) is independently hydrogen or(1-4C)alkyl; R⁴ represents a divalent hydrocarbon group containing from4 to 28 carbon atoms and optionally containing from 1 to 10 heteroatomsselected independently from halo, oxygen, nitrogen and sulfur; R⁵represents hydrogen or (1-4C)alkyl; R⁶ is —NR^(6a)CR^(6b)(O) or—CR⁶CR^(6d)OR^(6e) and R⁷ is hydrogen; or R⁶ and R⁷ together form—NR^(7a)C(O)CR^(7b)═CR^(7c), —CR^(7d)═CR^(7e)C(O)—NR^(7f)—,—NR^(7g)C(O)—CR^(7h)R^(7i)—CR^(7j)R^(7k)— or—CR^(7l)R^(7m)—CR^(7n)R^(7o)—C(O)—NR^(7p); each of R^(6a), R^(6b),R^(6c), R^(6d) and R^(6e) is independently hydrogen or (1-4C)alkyl; andeach of R^(7a), R^(7b), R^(7c), R^(7d), R^(7e), R⁷, R^(7g), R^(7h),R^(7i), R^(7j), R^(7k), R^(7l), R^(7m), R^(7n), R^(7o) and R^(7p) isindependently hydrogen or (1-4C)alkyl; or a stereoisomer thereof; theprocess comprising: (a) reacting a compound of formula 1:

or a salt thereof; with a compound of formula 2:

wherein X¹ represents a leaving group, and P¹ and P² each independentlyrepresent a hydrogen atom or a hydroxyl-protecting group; (b) reacting acompound of formula 3:

or salt thereof; with a compound of formula 4:

wherein X² represents a leaving group, and P³ and P⁴ each independentlyrepresent a hydrogen atom or a hydroxyl-protecting group; (c) coupling acompound of formula 5:

with a compound of formula 6:

wherein X^(Qa a)nd X^(Qb) each independently represent functional groupsthat couple to form a group Q, P^(5a) represents a hydrogen atom or anamino-protecting group; and P^(5b) and P⁶ each independently represent ahydrogen atom or a hydroxyl-protecting group; (d) for a compound offormula I wherein R⁵ represents a hydrogen atom, reacting a compound offormula 3 with a compound of formula 7:

or a hydrate thereof (e.g., a glyoxal), in the presence of a reducingagent, wherein P⁷ represents a hydrogen atom or a hydroxyl-protectinggroup; (e) reacting a compound of formula 1 with a compound of formula8:

or a hydrate thereof, in the presence of a reducing agent, wherein P⁸and P⁹ each independently represent a hydrogen atom or ahydroxyl-protecting group, P¹⁰ represents a hydrogen atom or anamino-protecting group, and R^(4′) represents a residue that, togetherwith the carbon to which it is attached, affords a group R⁴ uponcompletion of the reaction; (f) reacting a compound of formula 9:

wherein X³ represents a leaving group, with a compound of formula 10:

wherein P¹¹ and P¹² each independently represent a hydrogen atom or ahydroxyl-protecting group, and P¹³ represents a hydrogen atom or anamino-protecting group; or (g) reacting a compound of formula 11:

or a hydrate thereof; wherein R^(4′) represents a residue that, togetherwith the carbon to which it is attached, affords a group R⁴ uponcompletion of the reaction; with a compound of formula 10 in thepresence of a reducing agent; and (h) removing any protecting groups toform a compound of formula I; (i) repeating step (a), (b), (c), (d),(e), (f) or (g) and step (h) to prepare a library comprising compoundsof formula I.